Renal-homing peptide conjugates and methods of use thereof

ABSTRACT

Peptides that home, target, migrate to, are directed to, are retained by, or accumulate in and/or bind to the kidney of a subject are disclosed. Pharmaceutical compositions and uses for peptides or peptide-active agent complexes comprising such peptide-active agent conjugates are also disclosed. Such compositions can be formulated for targeted delivery of an active agent to a target region, tissue, structure or cell in the kidney. Targeted compositions of the disclosure can deliver peptide or peptide-active agent complexes to target regions, tissues, structures, or cells targeted by the peptide.

CROSS REFERENCE

This application is a U.S. National Phase Application under 35 U.S.C. §371 of International Application No. PCT/US2018/037544, filed Jun. 14,2018, which claims the benefit of U.S. Provisional Patent ApplicationNo. 62/520,323, filed Jun. 15, 2017, the entire disclosures of which areincorporated by reference.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Jun. 29, 2018, isnamed 45639-712_601_SL.txt and is 380,367 bytes in size.

BACKGROUND

Approximately 9% of the world's population either has, or is expected todevelop, chronic renal disease. The leading causes in the United Statesare diabetic nephropathy and progressive renal dysfunction following about of ischemic (e.g., post-cardiac surgery) or toxin-induced (e.g.,radiocontrast media, cancer chemotherapy) kidney proximal tubule damage.At present, the US End Stage Renal Disease (ESRD) program consumes ˜7%of the entire Medicare budget. Furthermore, even modest declines inrenal function can represent progressive, independent risk factors forrising hospital expenditures, morbidity and mortality. Thus, new ways toprotect kidneys, and prophylactically prevent and treat progressiverenal diseases are needed.

SUMMARY

The present disclosure relates to compositions and methods for treatmentof renal disorders. Described herein are peptides that home to, migrateto, accumulate in, bind to, are retained by, or are directed to, and/orbind in kidney following administration in a subject. In someembodiments, the homing peptides of the present disclosure are used todeliver a detection agent to image and/or diagnose renal injury, ordisease. In other embodiments, the homing peptides of the presentdisclosure are used to treat or deliver an active agent to a region,tissue, structure, or cell thereof.

In some aspects, a peptide active agent conjugate comprises a) apeptide, wherein the peptide comprises a sequence that has at least 70%sequence identity with any one of SEQ ID NO: 236-SEQ ID NO: 276, whereinupon administration to a subject the peptide homes, targets, migratesto, accumulates in, binds to, is retained by, or is directed to a kidneyof the subject; and an active agent selected from an active agent classselected from TABLE 5 or TABLE 6; b) a peptide, wherein the peptidecomprises a sequence that has at least 70% sequence identity with anyone of SEQ ID NO: 1-SEQ ID NO: 41, wherein upon administration to asubject the peptide homes, targets, migrates to, accumulates in, bindsto, is retained by, or is directed to a kidney of the subject; and anactive agent selected from TABLE 5 or TABLE 6; c) a peptide, wherein thepeptide comprises a sequence that has at least 70% sequence identitywith any one of SEQ ID NO: 471-SEQ ID NO: 529, wherein uponadministration to a subject the peptide homes, targets, migrates to,accumulates in, binds to, is retained by, or is directed to a kidney ofthe subject; and an active agent selected from an active agent classselected from TABLE 5 or TABLE 6; d) a peptide, wherein the peptidecomprises a sequence that has at least 70% sequence identity with anyone of SEQ ID NO: 277-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 470, wherein uponadministration to a subject the peptide homes, targets, migrates to,accumulates in, binds to, is retained by, or is directed to a kidney ofthe subject; and an active agent selected from an active agent classselected from TABLE 6; or e) a peptide, wherein the peptide comprises asequence that has at least 70% sequence identity with any one of SEQ IDNO: 42-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, orSEQ ID NO: 216-SEQ ID NO: 235, wherein upon administration to a subjectthe peptide homes, targets, migrates to, accumulates in, binds to, isretained by, or is directed to a kidney of the subject; and an activeagent selected from an active agent class selected from TABLE 6. Invarious aspects, the active agent is selected from TABLE 5.

In some aspects, the peptide active agent conjugate homes, targets,migrates to, accumulates in, binds to, is retained by, or is directed toa kidney of the subject. In other aspects, the peptide homes, targets,migrates to, accumulates in, binds to, is retained by, or is directed toproximal tubules of the kidney.

In various aspects, the peptide comprises: a) a sequence that has atleast 75%, at least 80%, at least 85%, at least 90%, or at least 95%, atleast 97%, at least 99% or 100% sequence identity with any one of SEQ IDNO: 236-SEQ ID NO: 276, or a fragment thereof; b) a sequence that has atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, atleast 97%, at least 99%, or 100% sequence identity with any one of SEQID NO: 1-SEQ ID NO: 41 or a fragment thereof; c) a sequence that has atleast 75%, at least 80%, at least 85%, at least 90%, or at least 95%, atleast 97%, at least 99% or 100% sequence identity with any one of SEQ IDNO: 471-SEQ ID NO: 529 or a fragment thereof; d) a sequence that has atleast 75%, at least 80%, at least 85%, at least 90%, or at least 95%, atleast 97%, at least 99% or 100% sequence identity with any one of SEQ IDNO: 277-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448,or SEQ ID NO: 451-SEQ ID NO: 470 or a fragment thereof; or e) a sequencethat has at least 75%, at least 80%, at least 85%, at least 90%, or atleast 95%, at least 97%, at least 99% or 100% sequence identity with anyone of SEQ ID NO: 42-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQID NO: 213, or SEQ ID NO: 216-SEQ ID NO: 235 or a fragment thereof. Inother aspects, the peptide comprises: a) a sequence of any one of SEQ IDNO: 236-SEQ ID NO: 276 or a fragment thereof; b) a sequence of any oneof SEQ ID NO: 1-SEQ ID NO: 41 or a fragment thereof; c) a sequence ofany one of SEQ ID NO: 471-SEQ ID NO: 529 or a fragment thereof; d) asequence of any one of SEQ ID NO: 277-SEQ ID NO: 355, SEQ ID NO: 362-SEQID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 470 or afragment thereof; or e) a sequence of any one of SEQ ID NO: 42-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, or SEQ ID NO:216-SEQ ID NO: 235 or a fragment thereof. In various aspects, thepeptide comprises a) a sequence of any one of SEQ ID NO: 550-SEQ ID NO:569 or a fragment thereof; or b) a sequence of any one of SEQ ID NO:530-SEQ ID NO: 549 or SEQ ID NO: 570, or a fragment thereof.

In some aspects, the peptide is at least 30%, 40%, 50%, 60%, 70%, 80%,90%, 95%, 97%, 99%, or 100% identical to SEQ ID NO: 135. In someaspects, the peptide is at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%,97%, 99%, or 100% identical to SEQ ID NO: 42. In other aspects, thepeptide is at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or100% identical to SEQ ID NO: 45. In still other aspects, the peptide isat least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or 100%identical to SEQ ID NO: 217. In some aspects, the peptide is at least30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or 100% identical toSEQ ID NO: 48. In some aspects, the peptide is at least 30%, 40%, 50%,60%, 70%, 80%, 90%, 95%, 97%, 99%, or 100% identical to SEQ ID NO: 132.In other aspects, the peptide is at least 30%, 40%, 50%, 60%, 70%, 80%,90%, 95%, 97%, 99%, or 100% identical to SEQ ID NO: 54. In some aspects,the peptide is at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%,99%, or 100% identical to SEQ ID NO: 231. In some aspects, the peptideis at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or 100%identical to SEQ ID NO: 43. In other aspects, the peptide is at least30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or 100% identical toSEQ ID NO: 130. In some aspects, the peptide is at least 30%, 40%, 50%,60%, 70%, 80%, 90%, 95%, 97%, 99%, or 100% identical to SEQ ID NO: 44.In some aspects, the peptide is at least 30%, 40%, 50%, 60%, 70%, 80%,90%, 95%, 97%, 99%, or 100% identical to SEQ ID NO: 219. In someaspects, the peptide is at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%,97%, 99%, or 100% identical to SEQ ID NO: 131. In some aspects, thepeptide is at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or100% identical to SEQ ID NO: 33. In other aspects, the peptide is atleast 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or 100%identical to SEQ ID NO: 4. In some aspects, the peptide is at least 30%,40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or 100% identical to SEQ IDNO: 41. In some aspects the peptide is at least 30%, 40%, 50%, 60%, 70%,80%, 90%, 95%, 97%, 99%, or 100% identical to SEQ ID NO: 5. In someaspects, the peptide is at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%,97%, 99%, or 100% identical to SEQ ID NO: 6. In other aspects, thepeptide is at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or100% identical to SEQ ID NO: 196.

In some aspects, the peptide is covalently conjugated to the activeagent. In some aspects, the peptide active agent conjugate homes,targets, migrates to, accumulates in, binds to, is retained by, or isdirected to a kidney of the subject. In some aspects, the peptidecomprises 4 or more cysteine residues. In further aspects, the peptidecomprises three or more disulfide bridges formed between cysteineresidues, wherein one of the disulfide bridges passes through a loopformed by two other disulfide bridges. In some aspects, the peptidecomprises a plurality of disulfide bridges formed between cysteineresidues. In some aspects, the peptide comprises a disulfide through adisulfide knot.

In some aspects, at least one amino acid residue of the peptide is in anL configuration or, wherein at least one amino acid residue of thepeptide is in a D configuration. In some aspects, the sequence comprisesat least 11, at least 12, at least 13, at least 14, at least 15, atleast 16, at least 17, at least 18, at least 19, at least 20, at least21, at least 22, at least 23, at least 24, at least 25, at least 26, atleast 27, at least 28, at least 29, at least 30, at least 31, at least32, at least 33, at least 34, at least 35, at least 36, at least 37, atleast 38, at least 39, at least 40, at least 41, at least 42, at least43, at least 44, at least 45, at least 46, at least 47, at least 48, atleast 49, at least 50, at least 51, at least 52, at least 53, at least54, at least 55, at least 56, at least 57, at least 58 residues, atleast 59, at least 60, at least 61, at least 62, at least 63, at least64, at least 65, at least 66, at least 67, at least 68, at least 69, atleast 70, at least 71, at least 72, at least 73, at least 74, at least75, at least 76, at least 77, at least 78, at least 79, at least 80, orat least 81 residues.

In some aspects, any one or more K residues are replaced by an A or Rresidue or wherein any one or more A or R residues are replaced by for aK residue. In some aspects, any one or more M residues are replaced byany one of the I, L, or V residues. In some aspects, any one or more Lresidues are replaced by any one of the V, I, or M residues. In someaspects, any one or more I residues are replaced by any of the M, L, orV residues.

In other aspects, any one or more V residues are replaced by any of theM, I, or L residues. In still other aspects, any one or more G residuesare replaced by an A residue or wherein any one or more A residues arereplaced by a G residue. In some aspects, any one or more S residues arereplaced by a T residue or wherein any one or more T residues arereplaced by for an S residue. In some aspects, any one or more Qresidues are replaced by an N residue or wherein any one or more Nresidues are replaced by a Q residue. In some aspects, any one or more Dresidues are replaced by an E residue or wherein any one or more Eresidues are replaced by a D residue.

In some aspects, the peptide has a charge distribution comprising anacidic region and a basic region. In further aspects, the acidic regionis a nub. In other aspects, the basic region is a patch. In someaspects, the peptide comprises 5-12 basic residues. In some aspects, thepeptide comprises 0-5 acidic residues. In some aspects, the peptidecomprises 6 or more basic residues and 2 or fewer acidic residues. Insome aspects, the peptide comprises a 4-19 amino acid residue fragmentcontaining at least 2 cysteine residues, and at least 2 positivelycharged amino acid residues.

In other aspects, the peptide comprises a 20-70 amino acid residuefragment containing at least 2 cysteine residues, no more than 2 basicresidues and at least 2 positively charged amino acid residues. In stillother aspects, the peptide comprises at least 3 positively charged aminoacid residues. In some aspects, the positively charged amino acidresidues are selected from K, R, or a combination thereof.

In some aspects, the peptide has a charge greater than 2 atphysiological pH. In other aspects, the peptide has a charge greaterthan 3.5 at physiological pH. In still other aspects, the peptide has acharge greater than 4.5 at physiological pH. In some aspects, thepeptide has a charge greater than 5.5 at physiological pH. In otheraspects, the peptide has a charge greater than 6.5 at physiological pH.In other aspects, the peptide has a charge greater than 7.5 atphysiological pH. In still other aspects, the peptide has a chargegreater than 8.5 at physiological pH. In other aspects, the peptide hasa charge greater than 9.5 at physiological pH.

In some aspects, the peptide is selected from a potassium channelagonist, a potassium channel antagonist, a portion of a potassiumchannel, a sodium channel agonist, a sodium channel antagonist, acalcium channel agonist, a calcium channel antagonist, a hadrucalcin, atheraphotoxin, a huwentoxin, a kaliotoxin, a cobatoxin, or a lectin.

In further aspects, the lectin is SHL-Ib2. In some aspects, the peptideis arranged in a multimeric structure with at least one other peptide.

In further aspects, the multimeric structure comprises a dimer, trimer,tetramer, pentamer, hexamer, or heptamer. In some aspects, at least oneresidue of the peptide comprises a chemical modification. In furtheraspects, the chemical modification is blocking the N-terminus of thepeptide. In still further aspects, the chemical modification ismethylation, acetylation, or acylation. In other aspects, the chemicalmodification is: methylation of one or more lysine residues or analoguethereof; methylation of the N-terminus; or methylation of one or morelysine residue or analogue thereof and methylation of the N-terminus. Insome aspects, the peptide is linked to an acyl adduct.

In some aspects, the peptide is linked to an active agent. In furtheraspects, the active agent is fused with the peptide at an N-terminus ora C-terminus of the peptide. In some aspects, the active agent isanother peptide. In some aspects, the active agent is an antibody. Inother aspects, the active agent is an Fc domain, Fab domain, scFv, or Fvfragment. In still other aspects, the peptide fused with an Fc domaincomprises a contiguous sequence.

In further aspects, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 active agents arelinked to the peptide. In still further aspects, the peptide is linkedto the active agent via a cleavable linker. In some aspects, the peptideis linked to the active agent at an N-terminus, at the epsilon amine ofan internal lysine residue, at the carboxylic acid of an aspartic acidor glutamic acid residue, or a C-terminus of the peptide by a linker. Insome aspects, the peptide further comprises a non-natural amino acid,wherein the non-natural amino acid is an insertion, appendage, orsubstitution for another amino acid.

In some aspects, the peptide is linked to the active agent at thenon-natural amino acid by a linker. In some aspects, the linkercomprises an amide bond, an ester bond, a carbamate bond, a carbonatebond, a hydrazone bond, an oxime bond, a disulfide bond, a thioesterbond, a thioether bond, a triazole, a carbon-carbon bond, or acarbon-nitrogen bond. In further aspects, the cleavable linker comprisesa cleavage site for matrix metalloproteinases, thrombin, cathepsins, orbeta-glucuronidase. In other aspects, the linker is a hydrolyticallylabile linker. In other aspects, the linker is pH sensitive, reducible,glutathione-sensitive, or protease cleavable. In still other aspects,the peptide is linked to the active agent via a stable linker.

In some aspects, the peptide is linked to a detectable agent. In furtheraspects, the detectable agent is fused with the peptide at an N-terminusor a C-terminus of the peptide. In still further aspects, 1, 2, 3, 4, 5,6, 7, 8, 9, or 10 detectable agents are linked to the peptide. In someaspects, the peptide is linked to the detectable agent via a cleavablelinker.

In some aspects, the peptide is linked to the detectable agent at anN-terminus, at the epsilon amine of an internal lysine residue, or aC-terminus of the peptide by a linker. In further aspects, the peptidefurther comprises a non-natural amino acid, wherein the non-naturalamino acid is an insertion, appendage, or substitution for another aminoacid. In still further aspects, the peptide is linked to the activeagent at the non-natural amino acid by a linker.

In still further aspects, the linker comprises an amide bond, an esterbond, a carbamate bond, a hydrazone bond, an oxime bond, or acarbon-nitrogen bond. In some aspects, the cleavable linker comprises acleavage site for matrix metalloproteinases, thrombin, cathepsins, orbeta-glucuronidase. In other aspects, the peptide is linked to thedetectable agent via a stable linker.

In some aspects, the detectable agent is a fluorophore, a near-infrareddye, a contrast agent, a nanoparticle, a metal-containing nanoparticle,a metal chelate, an X-ray contrast agent, a PET agent, a radioisotope,or a radionuclide chelator. In further aspects, the detectable agent isa fluorescent dye. In some aspects, the peptide has an isoelectric pointof about 9.

In some aspects, the peptide is SEQ ID NO: 45. In other aspects, thepeptide is SEQ ID NO: 132. In still other aspects, the peptide is SEQ IDNO: 33. In some aspects, the peptide is SEQ ID NO: 4. In some aspects,the peptide is SEQ ID NO: 41. In other aspects, the peptide is SEQ IDNO: 5. In still other aspects, the peptide is SEQ ID NO: 6. In someaspects, the peptide is SEQ ID NO: 570. In some aspects, the peptidecomprises at least 4, at least 5, at least 6, at least 7, at least 8, atleast 9, at least 10, at least 11, at least 12, at least 13, at least14, at least 15, or at least 16 cysteine residues.

In some aspects, at least one amino acid residue of the peptide is in anL configuration or, wherein at least one amino acid residue is in a Dconfiguration. In some aspects, the peptide comprises or is derived fromthe group consisting of: chlorotoxins, brazzeins, circulins, stecrisps,hanatoxins, midkines, hefutoxins, potato carboxypeptidase inhibitors,bubble proteins, attractins, αGI, α-GID, μpIIIA, ω-MVIIA, ω-CVID,χ-MrIA, ρ-TIA, conantokin G, contulakin G, GsMTx4, margatoxins, shK,toxin K, chymotrypsin inhibitors (CTI), EGF epiregulin core,hainantoxins, theraphotoxins, hexatoxins, opicalcins, imperatoxins,defensins, and insectotoxins. In some aspects, the peptide comprises oris derived from a human protein or peptide.

In some aspects, the peptide comprises an isoelectric point less than orequal to about 7.5. In some aspects, the peptide comprises anisoelectric point greater than or equal to about 7.5. In other aspects,the peptide comprises an isoelectric point within a range from about 3.0to about 10.0. In some aspects, the peptide comprises a non-uniformcharge distribution. In some aspects, the peptide comprises one or moreregions of concentrated positive charge. In some aspects, the peptidecomprises one or more regions of concentrated negative charge.

In some aspects, the composition comprises a mass-average molecularweight (Mw) less than or equal to 6 kDa, less than or equal to about 50kDa, or less than or equal to about 60 kDa. In some aspects, thecomposition comprises a mass-average molecular weight (Mw) within arange from about 0.5 kDa to about 50 kDa, or within a range from about0.5 kDa to about 60 kDa. In some aspects, the peptide is stable at pHvalues greater than or equal to about 7.0. In some aspects, the peptideis stable at pH values less than or equal to about 5.0, less than orequal to about 3.0, or within a range from about 3.0 to about 5.0. Insome aspects, the peptide is stable at pH values within a range fromabout 5.0 to about 7.0. In some aspects, the peptide being stablecomprises one or more of: the peptide being capable of performing itstherapeutic effect, the peptide being soluble, the peptide beingresistant to protease degradation, the peptide being resistant toreduction, the peptide being resistant to pepsin degradation, thepeptide being resistant to trypsin degradation, the peptide beingreduction resistant, or the peptide being resistant to an elevatedtemperature.

In some aspects, upon administration to a subject, the peptide homes,targets, is directed to, accumulates in, migrates to, is retained by, orbinds to renal tissue of the subject. In further aspects, the peptidehomes, targets, is directed to, accumulates in, migrates to, is retainedby, or binds to one or more of: a cortex region, a glomerulus, aproximal tubule, a medulla region, a descending tubule, an ascendingtubule, a loop of Henle, or a Bowman's capsule of the subject. In stillfurther aspects, the peptide homes, targets, is directed to, accumulatesin, migrates to, is retained by, or binds to a proximal tubule of thesubject.

In further aspects, the peptide homes, targets, is directed to,accumulates in, migrates to, is retained by, or binds to a cell of theproximal tubule. In some aspects, the peptide homes, targets, isdirected to, accumulates in, migrates to, is retained by, or binds to acell surface receptor expressed by the cell of the proximal tubule. Insome aspects, the peptide homes, targets, is directed to, accumulatesin, migrates to, is retained by, or binds to a glomerulus of thesubject. In other aspects, the peptide homes, targets, is directed to,accumulates in, migrates to, is retained by, or binds to a megalinreceptor, a cubulin receptor, or a combination thereof.

In some aspects, the peptide is internalized by a cell. In furtheraspects, the peptide is internalized by the cell via a scavengingmechanism. In some aspects, the peptide exhibits a renal therapeuticeffect. In further aspects, the renal therapeutic effect comprises arenal protective effect or renal prophylactic effect. In some aspects,the peptide interacts with a renal ion channel, inhibits a protease, hasantimicrobial activity, has anticancer activity, has anti-inflammatoryactivity, induces ischemic preconditioning or acquired cytoresistance,or produces a protective or therapeutic effect on a kidney of thesubject, or a combination thereof.

In some aspects, the active agent comprises a renal therapeutic agent.In further aspects, the renal therapeutic agent accumulates in thekidney at a higher level when linked to the peptide than when not linkedto the peptide. In some aspects, the renal therapeutic agent comprises arenal protective agent or a renal prophylactic agent. In some aspects,the renal therapeutic agent, renal protective agent, or renalprophylactic agent induces ischemic preconditioning or acquiredcytoresistance in a kidney of the subject. In some aspects, the activeagent interacts with a renal ion channel, inhibits a protease, hasantimicrobial activity, has anticancer activity, has anti-inflammatoryactivity, has a diuretic effect, increases glucose excretion, modulatesthe immune system, induces ischemic preconditioning or acquiredcytoresistance, produces a protective or therapeutic effect on a kidneyof the subject, reduces a clearance rate of the composition, or acombination thereof.

In some aspects, the composition further comprises a half-life modifyingagent coupled to the peptide. In further aspects, the half-lifemodifying agent comprises a polymer, a polyethylene glycol (PEG), ahydroxyethyl starch, polyvinyl alcohol, a water soluble polymer, azwitterionic water soluble polymer, a water soluble poly(amino acid), awater soluble polymer of proline, alanine and serine, a water solublepolymer containing glycine, glutamic acid, and serine, an Fc region, afatty acid, palmitic acid, or a molecule that binds to albumin.

In some aspects, administration of the composition to a patient mediatesinflammation, cell death, fibrosis, or any combination thereof in thekidney. In some aspects, the peptide active agent complex is expressedas a fusion protein. In some aspects, the renal ion channel is a calciumchannel, a magnesium channel, a chlorine channel, a hydrogen channel, apotassium channel, a sodium channel, or any combination thereof.

In various aspects, the present disclosure provides a pharmaceuticalcomposition comprising any composition described above or a saltthereof, and a pharmaceutically acceptable carrier. In some aspects, thepharmaceutical composition is formulated for administration to asubject. In further aspects, the pharmaceutical composition isformulated for inhalation, intranasal administration, oraladministration, topical administration, parenteral administration,intravenous administration, subcutaneous administration, intramuscularadministration, intraperitoneal administration, dermal administration,transdermal administration, or a combination thereof.

In various aspects, the present disclosure provides a method of treatinga condition in a subject in need thereof, the method comprising:administering to the subject a peptide comprising any compositiondescribed above or any pharmaceutical composition described above. Insome aspects, the composition is administered by inhalation,intranasally, orally, topically, parenterally, intravenously,subcutaneously, intramuscularly administration, intraperitoneally,dermally, transdermally, or a combination thereof. In some aspects, thecomposition homes, targets, or migrates to a kidney of the subjectfollowing administration.

In some aspects, the condition is associated with a function of a kidneyof the subject. In some aspects, the composition or pharmaceuticalcomposition homes, targets, or migrates to renal tissue of the subjectfollowing administration. In some aspects, the condition is associatedwith a function of a kidney of the subject.

In further aspects, the condition is selected from the group consistingof: acute kidney diseases and disorders (AKD), acute kidney injury,acute and rapidly progressive glomerulonephritis, acute presentations ofnephrotic syndrome, acute pyelonephritis, acute renal failure,idiopathic chronic glomerulonephritis, secondary chronicglomerulonephritis, chronic heart failure, chronic interstitialnephritis, chronic kidney disease (CKD), chronic liver disease, chronicpyelonephritis, diabetes, diabetic kidney disease, fibrosis, focalsclerosis, focal segmental glomerulosclerosis, Goodpasture's disease,diabetic nephropathy, hereditary nephropathy, interstitial nephropathy,hypertensive nephrosclerosis, IgG4-related renal disease, interstitialinflammation, lupus nephritis, nephritic syndrome, partial obstructionof the urinary tract, polycystic kidney disease, progressive renaldisease, renal cell carcinoma, clear cell renal cell carcinoma,papillary renal cell carincoma, chromophobe renal cell carinoma, kidneycancer, transitional cell carcinoma, nephroblastoma, renal sarcoma,renal adenoma, oncocytoma, angiomyolipoma, renal fibrosis, kidneystones, hypertension, hypotension, disorders of sodium, water,acid-base, potassium, calcium, magnesium, or phosphate balance,infections, urinary tract infections, kidney failure, hematuria, renalcysts, uremia, shock, uretal obstruction, proteinuria, Fanconi'ssyndrome, Bartter's syndrome, chronic renal insufficiency, renalfibrosis, graft versus host disease after renal transplant, organtransplant rejection, and vasculitis.

In still further aspects, the condition is lupus nephritis, acute kidneyinjury (AKI), chronic kidney disease (CKD), hypertensive kidney damage,diabetic nephropathy, or renal fibrosis.

In various aspects, the present disclosure provides a method of imagingan organ or body region of a subject, the method comprising:administering to the subject any composition described above or anypharmaceutical composition described above; and imaging the subject.

In some aspects, the method further comprises detecting a cancer ordiseased region, tissue, structure, or cell. In some aspects, the methodfurther comprises performing surgery on the subject. In further aspects,the method further comprises treating the cancer. In some aspects, thesurgery comprises removing the cancer or the diseased region, tissue,structure, or cell of the subject. In some aspects, the method furthercomprises imaging the cancer or diseased region, tissue, structure, orcell of the subject after surgical removal.

In various aspects, the present disclosure provides a method ofprotecting a kidney of a subject from injury, the method comprising:administering to the subject any composition described above or anypharmaceutical composition described above. In some aspects, thecomposition is administered by inhalation, intranasally, orally,topically, intravenously, subcutaneously, intramuscularlyadministration, intraperitoneally, or a combination thereof. In someaspects, the method further comprises inducing ischemic preconditioningor acquired cytoresistance in the kidney of the subject.

In some aspects, the injury is associated with one or more of: surgery,radiocontrast imaging, radiocontrast nephropathy, cardiovascularsurgery, cardiopulmonary bypass, extracorporeal membrane oxygenation(ECMO), balloon angioplasty, induced cardiac or cerebralischemic-reperfusion injury, organ transplantation, kidneytransplantation, sepsis, shock, low blood pressure, high blood pressure,kidney hypoperfusion, chemotherapy, drug administration, nephrotoxicdrug administration, blunt force trauma, puncture, poison, or smoking.

In some aspects, the composition or pharmaceutical composition isadministered at least 1 hour, at least 2 hours, at least 3 hours, atleast 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, atleast 8 hours, at least 9 hours, at least 10 hours, at least 11 hours,at least 12 hours, at least 13 hours, at least 14 hours, at least 15hours, at least 16 hours, at least 17 hours, at least 18 hours, at least19 hours, at least 20 hours, at least 21 hours, at least 22 hours, atleast 23 hours, at least 24 hours, at least 36 hours, at least 48 hours,at least 60 hours, at least 72 hours, or at least 96 hours prior to apredicted occurrence of the injury. In some aspects, the composition orpharmaceutical composition is administered once per day, week, or month,or once per two weeks, two months, or three months.

In some aspects, the composition or pharmaceutical composition isadministered at least 1 hour, at least 2 hours, at least 3 hours, atleast 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, atleast 8 hours, at least 9 hours, at least 10 hours, at least 11 hours,at least 12 hours, at least 13 hours, at least 14 hours, at least 15hours, at least 16 hours, at least 17 hours, at least 18 hours, at least19 hours, at least 20 hours, at least 21 hours, at least 22 hours, atleast 23 hours, at least 24 hours, at least 36 hours, at least 48 hours,at least 60 hours, at least 72 hours, or at least 96 hours after anoccurrence of the injury. In some aspects, the method further comprisesperforming a medical procedure on the subject.

In further aspects, the medical procedure comprises one or more of:surgery, radiocontrast imaging, cardiopulmonary bypass, balloonangioplasty, induced cardiac or cerebral ischemic-reperfusion injury,organ transplantation, chemotherapy, drug administration, or nephrotoxicdrug administration. In some aspects, the composition or thepharmaceutical composition is administered at least 1 hour, at least 2hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10hours, at least 11 hours, at least 12 hours, at least 13 hours, at least14 hours, at least 15 hours, at least 16 hours, at least 17 hours, atleast 18 hours, at least 19 hours, at least 20 hours, at least 21 hours,at least 22 hours, at least 23 hours, at least 24 hours, at least 36hours, at least 48 hours, at least 60 hours, at least 72 hours, or atleast 96 hours prior to performing the medical procedure.

In some aspects, the composition or the pharmaceutical composition isadministered at least 1 hour, at least 2 hours, at least 3 hours, atleast 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, atleast 8 hours, at least 9 hours, at least 10 hours, at least 11 hours,at least 12 hours, at least 13 hours, at least 14 hours, at least 15hours, at least 16 hours, at least 17 hours, at least 18 hours, at least19 hours, at least 20 hours, at least 21 hours, at least 22 hours, atleast 23 hours, at least 24 hours, at least 36 hours, at least 48 hours,at least 60 hours, at least 72 hours, or at least 96 hours afterperforming the medical procedure.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned, disclosedor referenced in this specification are herein incorporated by referencein their entirety and to the same extent as if each individualpublication, patent, or patent application was specifically andindividually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE FIGURES

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present disclosure will be obtained by reference tothe following detailed description that sets forth illustrativeembodiments, in which the principles of the disclosure are utilized, andthe accompanying drawings of which:

FIG. 1 illustrates an exemplary architecture of constructs expressingsequences of SEQ ID NO: X, where X can be any one of peptides of SEQ IDNO: 21-SEQ ID NO: 33.

FIG. 2 illustrates a schematic of a method of manufacturing of a peptideof the disclosure.

FIG. 3 shows renal signal patterns for a peptide of SEQ ID NO: 4. FIG.3A shows accumulation of ¹⁴C signal for radiolabeled SEQ ID NO: 4 threehours after peptide administration. FIG. 3B shows accumulation of ¹⁴Csignal for a peptide of SEQ ID NO: 4 twenty-four hours after peptideadministration.

FIG. 4 shows whole body fluorescence images of mice after administrationof SEQ ID NO: 132 conjugated to Cy5.5 (SEQ ID NO: 132-Cy5.5) (left)versus after administration of free Cy5.5—COOH alone (right). FIG. 4Ashows a whole body fluorescence image of a mouse 3 hours afteradministration of 10 nmol SEQ ID NO: 132-Cy5.5. The arrow indicates theposition and fluorescence signal in the kidney. FIG. 4B shows a wholebody fluorescence image of a mouse 3 hours after administration of 10nmol Cy5.5—COOH. The arrow indicates the position and fluorescencesignal in the kidney. FIG. 4C shows a whole body fluorescence image of amouse after 24 hours after administration of 10 nmol SEQ ID NO:132-Cy5.5. The arrow indicates the position and fluorescence signal inthe kidney. FIG. 4D shows a whole body fluorescence image of a mouse 24hours after administration of 10 nmol Cy5.5—COOH. The arrow indicatesthe position and fluorescence signal in the kidney. FIG. 4E shows awhole body fluorescence image of a mouse 48 hours after administrationof 10 nmol SEQ ID NO: 132-Cy5.5. The arrow indicates the position andfluorescence signal in the kidney. FIG. 4F shows a whole bodyfluorescence image of a mouse 48 hours after administration of 10 nmolCy5.5—COOH. The arrow indicates the position and fluorescence signal inthe kidney. FIG. 4G shows a whole body fluorescence image of a mouse 72hours after administration of 10 nmol SEQ ID NO: 132-Cy5.5. The arrowindicates the position and fluorescence signal in the kidney. FIG. 4Hshows a whole body fluorescence image of a mouse 72 hours afteradministration of 10 nmol Cy5.5—COOH. The arrow indicates the positionand fluorescence signal in the kidney.

FIG. 5 shows fluorescence of kidney sections from mice, in which eachmouse received 10 nmol free AlexFluor 647 fluorophore (AF647), 10 nmolSEQ ID NO: 41 conjugated to AF647, 10 nmol SEQ ID NO: 5 conjugated toAF647, or 10 nmol SEQ ID NO: 33 conjugated to AF647. Each kidney wasfrom an independent mouse.

FIG. 6 shows SEQ ID NO: 5 conjugated to AF647 and SEQ ID NO: 41conjugated to AF647 fluorescence signal in confocal images of the kidneycortex. FIG. 6A shows fluorescence signal of SEQ ID NO: 5 conjugated toAF647 in the kidney cortex 20 hours after of administration of 10 nmolof the peptide-dye conjugate at 6× magnification. FIG. 6B showsfluorescence signal of SEQ ID NO: 5 conjugated to AF647 in the kidneycortex 20 hours after administration of 10 nmol of the peptide-dyeconjugate at 20× magnification. FIG. 6C shows fluorescence signal of SEQID NO: 5 conjugated to AF647 in the kidney cortex 20 hours afteradministration of 10 nmol of the peptide-dye conjugate at 6×magnification. FIG. 6D shows fluorescence signal of SEQ ID NO: 5conjugated to AF647 in the kidney cortex 20 hours after ofadministration of 10 nmol of the peptide-dye conjugate at 20×magnification.

FIG. 7 shows SEQ ID NO: 33 conjugated to AF647 fluorescence signal inconfocal images of the kidney cortex. FIG. 7A shows fluorescence signalof SEQ ID NO: 33 conjugated to AF647 in the kidney cortex 20 hours afteradministration of 10 nmol of the peptide-dye conjugate at 6×magnification. FIG. 7B shows fluorescence signal of SEQ ID NO: 33conjugated to AF647 in the kidney cortex 20 hours after administrationof 10 nmol of the peptide-dye conjugate at 20× magnification. FIG. 7Cshows fluorescence signal in the kidney cortex 20 hours afteradministration of 10 nmol of a lysozyme-dye conjugate at 6×magnification. FIG. 7D shows fluorescence signal in the kidney cortex 20hours after of administration of 10 nmol of a lysozyme-dye conjugate at20× magnification.

FIG. 8 shows quantified fluorescence signal, indicating renal uptake, ofa peptide of SEQ ID NO: 4 conjugated to AlexaFluor647 (AF647) and anunlabeled SEQ ID NO: 4 peptide 4 hours after intravenous administrationof 2 nmol of SEQ ID NO: 4-AF647, 10 nmol of SEQ ID NO: 4 (1:5)co-injected with 2 nmol of SEQ ID NO: 4-AF647 (5:1), or 50 nmol of SEQID NO: 4 co-injected with 2 nmol of SEQ ID NO: 4-AF647 (25:1). Kidneysfrom uninjected mice were used as a negative control.

FIG. 9 shows quantified fluorescence signal, indicating renal uptake,between a peptide of SEQ ID NO: 4 conjugated to AlexaFluor647 (AF647)and unlabeled KKEEEKKEEEKKEEEKK competitor peptide (SEQ ID NO: 571, aknown renal targeting peptide) 1 hour after intravenous administrationof 2 nmol of a peptide of SEQ ID NO: 4-AF647, 2 nmol of a peptide of SEQID NO: 4-AF647 co-injected with 100 nmol of an unlabeled peptide of SEQID NO: 571 (1:50), or 2 nmol of peptide of SEQ ID NO: 4-AF647co-injected with 2000 nmol of an unlabeled peptide of SEQ ID NO: 571(1:1000).

FIG. 10 shows quantified fluorescence signal, indicating renal uptake,between a peptide of SEQ ID NO: 4 conjugated to AlexaFluor647 (AF647)and a control peptide conjugated to AF647 (control peptide-AF647), 4hours after intravenous administration of 10 nmol of a peptide of SEQ IDNO: 4-AF647 or 10 nmol of a peptide of control peptide-AF647.1

DETAILED DESCRIPTION

The present disclosure relates generally to compositions and methods forrenal therapy. In some embodiments, the compositions and methods hereinutilize peptides that home, target, are directed to, are retained by,accumulate in, migrate to, and/or bind to a kidney followingadministration to a subject. In some embodiments, the kidney homingpeptides of the present disclosure exert a therapeutic effect in akidney, or a tissue or a cell thereof. In some embodiments, the kidneyhoming peptides of the present disclosure are used to deliver an activeagent to a kidney, or a tissue or a cell thereof. The active agent canexert a therapeutic effect on a kidney, or a tissue or a cell thereof.For example, in certain embodiments, the peptide itself or the activeagent allows for localized delivery of an anti-inflammatory or otheragent to a kidney, or a tissue or a cell thereof. As another example,the active agent is a fluorophore that can be used for imaging of akidney. In certain embodiments, the peptide itself induces therapeuticresponses.

Kidney disorders can be particularly difficult to treat. A direct routefor active agent administration to the kidney can be parenterally (e.g.,intravenously, subcutaneously, intramuscularly), directly into thekidney, intra-articularly, by inhalation, dermally, topically, ororally. Drugs for kidney diseases can be injected directly locally intothe affected area, for example, directly injected into the kidney orspecific structures within the kidney. Few drugs aimed at treatingkidney disorders have proved therapeutically viable due primarily torapid clearance in a kidney as well as a lack of access to the targetkidney tissue. The lack of access to the target tissue and rapidclearance can also lead to administration of doses that are higher thanwould be necessary if a drug could home, target, or be directed to, isretained by, and/or binds to a target region, tissue, structure, orcell. Thus, treatment of kidney conditions often requires the use ofhigh concentrations of non-specific drugs. In addition, a number oftherapeutics are of interest in treating kidney disorders, but areproblematic because of the level of side effects caused by systemicadministration of the drug (Brenner and Rector's The Kidney, Skorecki etal, Elsevier, 10^(th) Edition, 2016).

Specific and potent drugs that are capable of contacting the kidney cancounteract the non-specificity of many treatments by selectivelytargeting and delivering compounds to specific regions, tissues, cellsand structures. Such drugs can also be useful to modulate ion channels,protein-protein interactions, extracellular matrix remodeling (e.g.,protease inhibition), and the like. Such targeted therapy can allow forlower dosing, reduced side effects, improved patient compliance, andimprovement in therapeutic outcomes, which would be advantageous notonly in acute disease of the kidney, but in chronic conditions as well.

The present disclosure provides peptides that can comprise or can bederived from cystine-dense peptides. As used herein, the term“cystine-dense peptide” can be interchangeable with the terms “knottedpeptide,” “knottin,” and “optide,” and cystine-dense peptides can alsobe abbreviated as “CDPs.” Hitchins, amongst other disulfide-containingpeptides, can also be considered “knotted peptides” or “cystine-densepeptides” for the purposes of this disclosure. Knottins, for example,are a class of cystine-dense peptides comprising from about 11 to about80 amino acids in length that are often folded into a compact structure.Knottins and other cystine-dense peptides are typically assembled into acomplex tertiary structure that is characterized by a number ofintramolecular disulfide crosslinks and can contain beta strands, analpha helix, and other secondary structures. The presence of thedisulfide bonds can give cystine-dense peptides remarkable environmentalstability, allowing them to withstand extremes of temperature and pH, toresist proteolytic enzymes in the blood stream or digestive tract, andcan provide specific biodistribution, pharmacokinetic, bindinginteractions, cellular processing, or other properties of physiologicand therapeutic value. The peptides disclosed herein can be derived fromcertain cystine-dense peptides. The present disclosure describes a classof cystine-dense peptides that can effectively contact kidney and beused either directly or as carriers of active drugs, peptides, ormolecules to treat a kidney condition.

Also described herein are peptides that can selectively home, target,are directed to, migrate to, are retained by, or accumulate in and/orbind to specific regions, tissues, structures, or cells of the kidneythat aid in managing, decreasing, ablating or reducing pain due tochronic disease or kidney injury or other therapeutic indications asdescribed herein. A peptide that homes, targets, migrates to, isdirected to, is retained by, or accumulates in and/or binds to one ormore specific regions, tissues, structures, or cells of the kidney canhave fewer off-target and potentially negative effects, for example,side effects that often limit use and efficacy of pain drugs. Inaddition, such peptides can reduce dosage and increase the efficacy ofexisting drugs by directly targeting them to a specific region, tissue,structure or cell of the kidney and helping the contact the kidney orincreasing the local concentration of agent. The peptide itself canmodulate pain or it can be conjugated to an agent that modulates pain.Such pain modulation may operate by various mechanisms such asmodulating inflammation, autoimmune responses, direct or indirect actionon pain receptors, cell killing, or programmed cell death (whether viaan apoptotic and/or non-apoptotic pathway of diseased cells or tissues,and the like (Tait et al., J Cell Sci 127(Pt 10):2135-44 (2014)).

Peptides of this disclosure that home, target, are directed to, migrateto, are retained by, accumulate in, or bind to specific regions,tissues, structures, or cells of the kidney can do so with differentdegrees of efficiency. Peptides can have a higher concentration inkidney than in other locations, such as blood or muscle. Peptides can berecorded as having a signal in kidney as a percentage of signal inblood. For example, a kidney signal of 200% indicates that the signal inkidney is twice as high as the signal in blood. In some embodiments,peptides that have kidney homing properties can have a kidney signal offrom >200% to >4000% by radiographic densitometry measurements. In otherembodiments, peptides that are kidney homers can have a kidney signalof >200% by radiographic densitometry measurements. In otherembodiments, peptides that are more efficient kidney homers can have akidney signal of >300% by radiographic densitometry measurements. Inother embodiments, peptides that are more efficient kidney homers canhave a kidney signal of >400% by radiographic densitometry measurements.In other embodiments, peptides that are strongest kidney homers ofhighest interest can have a kidney signal of >500% by radiographicdensitometry measurements, a kidney signal of >600% by radiographicdensitometry measurements, or a kidney signal of >4000% by radiographicdensitometry measurements. In some embodiments, peptides that are thestrongest kidney homers can have a kidney signal of from >600%to >4000%, from >600% to >700%, from >700% to >800%, from >800%to >900%, from >900% to >1000%, from >1000% to >2000%, from >2000%to >3000%, from >3000% to >4000%, or greater than 4000%. In someembodiments, measurement of the ratio of peptide concentration in blood,muscle, or other tissues relative to the peptide concentration in kidneycan be performed using various methods including measuring thedensitometry signal of peptides labeled with radioisotopes (as describedabove), or by using other assays.

Peptides that selectively home, target, are directed to, migrate to, areretained by, or accumulate in and/or bind to specific regions, tissues,structures, or cells of the kidney can occur after administration of thepeptide to a subject. A subject can be a human or a non-human animal.

The present disclosure relates generally to compositions and methods forrenal therapy. In some embodiments, the compositions and methods hereinutilize peptides that can home, target, are directed to, accumulate in,migrate to, are retained by and/or bind to the kidneys followingadministration to a subject. In certain embodiments, the peptidesdescribed herein can bind to or accumulate in a specific region, tissue,structure, or cell of a kidney, e.g., the proximal tubule, theglomerulus, or the glomerular filtrate (Bowman's space) tubular lumina.The properties of the peptide (e.g., isoelectric point (pI), molecularweight, pH stability, reduction resistance, protease resistance,hydrophobicity/hydrophilicity, charge, etc.) can be selected to provideimproved renal localization and binding. In some embodiments, the renalhoming peptides of the present disclosure are used to deliver an activeagent to the kidney or a tissue, region, compartment or cell thereof.The active agent can exert a therapeutic effect on the kidney or atissue or cell thereof. For example, in certain embodiments, the activeagent induces a protective response such as ischemic preconditioning oracquired cytoresistance in the kidney or tissue or cell thereof. Asanother example, in certain embodiments, the active agent induces atherapeutic response in a diseased kidney or tissue, region, compartmentor cell thereof. In certain embodiments, the peptide itself induces suchprotective and therapeutic responses, such as by binding to ionchannels, exerting an antimicrobial effect, or inhibiting protease(s).

Iron (Fe) mediated oxidative stress and renal interstitial inflammationcan lead to progressive nephron loss and renal interstitial fibrosis.The severity of the latter, as assessed on kidney biopsy, can be apredictor of subsequent loss of renal function. Despite recognition oftheir pathogenic roles, therapies targeted at Fe-mediated oxidativestress and renal inflammation have been hampered by two dominantfactors: 1) an inability to achieve sufficient intrarenal concentrationsof potent antioxidant/Fe binding agents (e.g., deferoxamine); and 2)associated systemic toxicities (e.g., with glucocorticoids,cyclophosphamide therapies). A molecule that can distribute sufficientlevels of a therapeutic agent to the kidney while reducing the levels ofthe agent delivered to other areas of the body such as to reduceoff-target toxicities may be able to achieve a therapeutic window thatallows treatment of the kidney with the agent with a sufficient safetyprofile. Likewise, an active molecule that can accumulate in the kidneywith reduced distribution to other tissues may be able to achieve atherapeutic effect in the kidney while sufficiently sparing othertissues from side effects. For example, steroid treatment of the kidneycan be limited by toxicity side effects in other parts of the body andin particular, can be contraindicated in diabetic patients due tooff-target toxicities.

In some embodiments, the present disclosure sets forth pro-drugs thatspecifically target the kidney. In some cases, low molecular weightproteins in plasma (LMWPs; <35 kDa) can be freely filtered by theglomerulus, and can be almost fully reabsorbed by proximal tubules(which represent ˜70% of total renal cortical mass). The reabsorbedprotein can be degraded within the proximal tubular lysosomal system.Thus, by binding small therapeutic molecules to a specific LMWP, thebound agent(s) can be tunably released from its carrier protein withintubular cells, gaining access to the tubular cytosol, and subsequently,the renal interstitial compartment (the dominant site of the renalinflammatory response).

The present disclosure provides a number of peptides that can berapidly, highly, and persistently taken up by or can accumulate inproximal tubule cells or in the glomerular filtrate (Bowman's space)tubular lumina. These peptides can prevent and treat a host of acute andprogressive renal diseases or can be linked to a therapeutic moleculethat can prevent and treat a host of acute and progressive renaldiseases. Given that many renal diseases, both acute and chronic, can bemediated in large part by both inflammation and iron mediated oxidativestress, the peptide-drug conjugates of the present disclosure can beapplicable in a wide range of clinical settings.

The peptides disclosed herein also can provide several advantages overother known approaches for treatment of acute or progressive renaldisease. For example, a peptide of this disclosure can deliver moleculesintracellularly, and thus act on intracellular targets as compared toother approaches. Additionally, as compared to treatment using lysozymeor myoglobin, a peptide of the disclosure can have reducedimmunogenicity, be soluble in kidney compartments, have a lack oftoxicity or reduced toxicity to kidney, and can be resistant toreduction and/or to proteases (Zhou et al. Acta Pharm Sin B. 2014February; 4(1):37-42). A peptide as disclosed herein can also have acontrolled and/or single site for drug conjugation as compared to otherknown treatments. For example, both a lysozyme (Haas 1997) and anotherpreviously known kidney targeting peptide, KKEEEKKEEEKKEEEKK (SEQ ID NO:571), can comprise multiple lysine residues as compared with a peptideof the disclosure, such as SEQ ID NO: 41, SEQ ID NO: 132, SEQ ID NO:130, SEQ ID NO: 131, SEQ ID NO: 135, SEQ ID NO: 219, SEQ ID NO: 4, SEQID NO: 5, and SEQ ID NO: 33, which have been or can be engineered tohave no lysine residue. The absence of a lysine residue on a peptide ofthe disclosure can allow for site specific amine conjugation at theN-terminus of the peptide or can allow for a single lysine residue to bea site specific conjugation. Lysine residues in some peptides can beessential for accumulation in the kidney, such that multipleconjugations on the lysine residues can reduce kidney accumulation andsubstitutions of lysine residues with arginine residues, to maintainpositive charge, can, with some peptides, result in reduced accumulationin kidney (Wischnjow et al. Bioconjug Chem. 2016 Apr. 20; 27(4):1050-7,Janzer et al. Bioconjug Chem. 2016 Oct. 4), whereas peptides of thisdisclosure can contain no lysine residues and still accumulate in thekidney.

The presence of multiple lysines on a peptide can result in multiplesites of conjugation, and can thereby result in heterogeneousconjugates. Similarly, therapeutic molecules like chitosan orpolyvinylpyrrolidone can also have multiple conjugation sites that canconfound preparation of a desired homogenous peptide drug conjugate (He2012, Wischnjow et al. Bioconjug Chem. 2016 Apr. 20; 27(4):1050-7).Furthermore, lysozyme can have cardiovascular side effects in comparisonwith a peptide of this disclosure, While small molecule drugs canreadily perfuse the kidney, a method for renal targeting can providedrug uptake into cells, accumulation of drug in the proximal tubules,retention of drug in the kidney, and can reduce systemic exposure to thedrug (Janzer et al. Bioconjug Chem. 2016 Oct. 4).

In some embodiments, any peptide of this disclosure can be grafted toanother moiety to enhance binding and/or accumulation in the kidney. Forexample, other targeting peptides can be grafted to any of the peptidesof this disclosure in order to enhance, change, or modify the propertiesof the peptides of the present disclosure. These other targetingpeptides can have positively charged residues, which can increasebinding of peptides to proximal tubule cells, to megalin (which isnegatively charged), or can otherwise increase retention in the kidney(Janzer et al. Bioconjug Chem. Oct. 4, 2016, Geng et al. Bioconjug Chem.Jun. 20, 2012; 23(6):1200-10, Wischnjow et al. Bioconjug Chem. Apr. 20,2016; 27(4):1050-7). Any of the peptide sequences described in Wischnjowet al. Bioconjug Chem. Apr. 20, 2016; 27(4):1050-7; Janzer et al.Bioconjug Chem. Oct. 4, 2016, Geng et al. Bioconjug Chem. Jun. 20, 2012;23(6):1200-10 can be grafted to a peptide of the present disclosure, andare incorporated herein by reference. These other peptides can modifythe properties of the peptides of the present disclosure by changingcharge, changing absorption properties into the proximal tubules, orchanging targeting of specific structures within the kidney.

For example, other targeting peptides can include Y(KKEEE)₃K (SEQ ID NO:624), Y(KKEE)₅K (SEQ ID NO: 625), Y(KKQQQ)₃K (SEQ ID NO: 626), Y(MARIA)₃(SEQ ID NO: 627), (KKEEE)₃K (SEQ ID NO: 628), (KKEE)₅K (SEQ ID NO: 629),(KKQQQ)₃K (SEQ ID NO: 630), (MARIA)₃ (SEQ ID NO: 631), (APASLYN)₂ (SEQID NO: 632), and ANTPCGPYTHDCPCKR (SEQ ID NO: 633). Any L-Tyr residue inany of the foregoing can be modified to D-Tyr, for example, for thepurposes of radiolabeling.

The peptides disclosed herein can be used as active agents, conjugatedto detection agents such a fluorophores, iodide-containing X-raycontrast agents, lanthanide chelates (e.g., gadolinium for MRI imaging),perfluorocarbons (for ultrasound), or PET tracers (e.g., 18F or 11C) forimaging and tracing the peptide, or conjugated to agents such asanti-inflammatory active agents or other active agents to the joint totreat inflammation or other disease.

The peptides disclosed herein can be used to bind kidney explants exvivo as well as kidney tissues, cells, and cell lines. Kidney explantscan be from any subject, such as a human or an animal. Assessment ofpeptide binding to kidney explants can be used to screen peptides thatmay efficiently home to kidney in vivo.

In some embodiments, peptides of this disclosure home, target, aredirected to, migrate to, are retained by, accumulate in, or bind tospecific regions, tissues, structures, or cells of the kidneys. Forexample, in some embodiments, peptides of this disclosure home, target,are directed to, migrate to, are retained by, accumulate in, or bind tothe proximal tubules of the kidneys, kidney nephrons, or podocytes.Peptides that selectively home, target, are directed to, migrate to, areretained by, or accumulate in and/or bind to specific regions, tissues,structures, or cells of the kidney can occur after administration of thepeptide to a subject. A subject can be a human or a non-human animal.The peptides disclosed herein can be used as active agents, orconjugated to detection agents such a fluorophores, iodide-containingX-ray contrast agents, lanthanide chelates (e.g., gadolinium for MRIimaging), perfluorocarbons (for ultrasound), or PET tracers (e.g. 18F or11C) for imaging and tracing the peptide, or conjugated to agents suchas anti-inflammatory agents or other agents to the kidney to treat renalcancer, chronic kidney failure or other kidney disease.

One roadblock in the advancement and wide spread use of peptides as atherapeutic is that peptides can be chemically and physically unstable.During the process of manufacturing of therapeutic peptides essentialconsiderations can include storage conditions, sustained biochemicalfunction, and in vivo delivery. Peptide degradation products can resultin the formation of species that alter the safety profile, potency, andimmunogenicity of the peptide. These peptide degradation products canform during manufacture and storage, as well as in vivo after deliveryto a patient. Furthermore, peptide degradation may limit the shelf-lifeand increase production cost due to unstable peptides requiringrefrigeration or shipment on dry ice. The latter can necessitatecontinual monitoring and validation of peptides as degradation productscould have formed during the manufacturing process. Hence, there is anurgent need for the rationale design and production of therapeuticpeptides that have enhanced stability, for example, in the ambientenvironment, during the process of manufacturing, in storage, and thatprevent the likelihood of peptide degradation under a variety ofconditions.

In some embodiments, the peptides and peptide-drug conjugates of thepresent disclosure have stability properties that minimize peptide orpeptide-drug conjugate degradation to enable adequate storage. Longterm, accelerated, and intermediate storage conditions for the peptidesand peptide-drug conjugates of the present disclosure can include longterm storage conditions of 25° C.±2° C./60% relative humidity (RH)±5%RH, or 30° C.±2° C./65% RH±5% RH for at least 6 months, at least 12months, and up to 1 year, up to 2 years, up to 3 years, up to 4 years,or longer than 4 years. In addition, intermediate and short term storageconditions (e.g., during transport, distribution, manufacturing, orhandling), or long term storage conditions for certain climates andinfrastructures, can include storage conditions of 30° C.±2° C./65%RH±5% RH or 40° C.±2° C./75% RH±5% RH for up to 1 hour, for up to 8hours, for up to 1 day, for up to 3 days, for up to 1 week, for up to 1month, for up to 3 months, for up to 6 months or at least 6 months, upto 1 year, up to 2 years, up to 3 years, up to 4 years, or longer than 4years. Moreover, the peptides and peptide-drug conjugates of the presentdisclosure can be refrigerated, for example between 5° C.±3° C. for atleast 6 months, at least 12 months, and up to 1 year, up to 2 years, upto 3 years, up to 4 years, or longer than 4 years. In addition,intermediate and short term refrigeration conditions (e.g., duringtransport, distribution, manufacturing, or handling) can include 25°C.±2° C./60% RH±5% RH for up to 1 hour, for up to 8 hours, for up to 1day, for up to 3 days, for up to 1 week, for up to 1 month, for up to 3months, for up to 6 months or at least 6 months, and potentially longer(at least 12 months and up to 1 year, up to 2 years, up to 3 years, upto 4 years, or longer than 4 years). Such conditions for storage,whether based on ambient or refrigerated conditions can be adjustedbased upon the four zones in the world (e.g., the International Councilfor Harmonisation of Technical Requirements for Pharmaceuticals forHuman Use (ICH) stability Zone I, II, III, or IV) that are distinguishedby their characteristic prevalent annual climatic conditions. Inaddition, formulation components can be principally chosen for theirability to preserve the native conformation and chemical structure ofthe peptides and peptide-drug conjugates of the present disclosure instorage by preventing denaturation due to hydrophobic interactions andaggregation, as well as by preventing chemical degradation, includingtruncation, oxidation, deamidation, cleavage, hydrolysis, isomerization,disulfide exchange, racemization, and beta elimination (Cleland, et al.,Crit Rev Ther Drug Carrier Syst 10(4): 307-377 (1993); Shire et al., JPharm Sci 93(6): 1390-1402 (2004); Wakankar and Borchardt, J Pharm Sci95(11): 2321-2336 (2006)).

In some embodiments, the peptides and peptide-drug conjugates of thepresent disclosure have incorporated properties that minimizeimmunogenicity of the peptides and peptide-drug conjugates.Immunogenicity can be a major concern with the development oftherapeutic peptides and proteins, and there is an urgent need for therationale design and production of therapeutic peptides that havereduced immunogenicity and that increase their safety and efficacy.Immunogenicity can occur against a desired peptide sequence or a peptidedegradation product. Immunogenicity can occur when a patient develops animmune response to the therapeutic peptide, protein, conjugate, or otherdrug, such as by producing antibodies that bind to and/or neutralize thetherapeutic peptide, protein, conjugate, or other drug. The likelihoodof immunogenicity can increase when drugs are administered more thanonce or chronically. Immunogenicity can reduce patient exposure to thedrug, can reduce effectiveness of the drug, and can also result insafety risks for the patient, such as generating an immune response toself-proteins or other adverse responses related to increasedimmunogenicity to the therapeutic peptide, protein, conjugate, or otherdrug. Immunogenic responses can vary from patient to patient and alsoamongst different groups of HLA alleles, as well as over time. As such,minimizing risk of immunogenicity with a therapeutic peptide or proteincan be important for developing a drug that can be effectively andsafely used for treatment. Various methods exist for assessment ofimmunogenic potential, which can include in silico methods, in vitrotesting, preclinical in vivo testing, and assessment during clinicaldosing. Evaluation early in product design and development of thetherapeutic peptides and peptide-drug conjugates of the presentdisclosure in the in vivo milieu in which they function (e.g., ininflammatory environments or at physiologic pH) can revealsusceptibilities to modifications (e.g., aggregation and deamidation)that can result in loss of efficacy or induction of immune responses.Such information can be used to facilitate product engineering toenhance the stability of the product under such in vivo conditions orreduce immunogenicity. Moreover, the therapeutic peptides andpeptide-drug conjugates of the present disclosure can be designed tominimize protein aggregation. Strategies to minimize aggregate formationcan be used early in drug development, for example, by using anappropriate cell substrate, selecting manufacturing conditions thatminimize aggregate formation, employing a robust purification schemethat removes aggregates to the greatest extent possible, and choosing aformulation and container closure system that minimize aggregationduring storage.

Additional aspects and advantages of the present disclosure will becomeapparent to those skilled in this art from the following detaileddescription, wherein illustrative embodiments of the present disclosureare shown and described. As will be realized, the present disclosure iscapable of other and different embodiments, and its several details arecapable of modifications in various respects, all without departing fromthe disclosure. Accordingly, the drawings and description are to beregarded as illustrative in nature, and not as restrictive.

As used herein, the abbreviations for the natural L-enantiomeric aminoacids are conventional and are as follows: alanine (A, Ala); arginine(R, Arg); asparagine (N, Asn); aspartic acid (D, Asp); cysteine (C,Cys); glutamic acid (E, Glu); glutamine (Q, Gln); glycine (G, Gly);histidine (H, His); isoleucine (I, Ile); leucine (L, Leu); lysine (K,Lys); methionine (M, Met); phenylalanine (F, Phe); proline (P, Pro);serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y,Tyr); valine (V, Val). Typically, Xaa can indicate any amino acid. Insome embodiments, X can be asparagine (N), glutamine (Q), histidine (H),lysine (K), or arginine (R). D amino acids are denoted with lower caseletters.

Some embodiments of the disclosure contemplate D-amino acid residues ofany standard or non-standard amino acid or analogue thereof. When anamino acid sequence is represented as a series of three-letter orone-letter amino acid abbreviations, the left-hand direction is theamino terminal direction and the right-hand direction is the carboxyterminal direction, in accordance with standard usage and convention.

Peptides

The cystine-dense peptides herein can bind targets with antibody-likeaffinity. The cystine-dense peptides can modulate the activity of aplurality of renal regions, tissues, structures, or cells. For example,in some embodiments, the cystine-dense peptide conjugated to achemotherapeutic or pain-modifying drug homes to the kidney of adiseased kidney and releases the drug, creating a higher localconcentration of drug in an area of diseased or damaged kidney thanwould be achieved without the kidney targeting function of the peptide.The cystine-dense peptide can be conjugated to a drug that can affectnearby tissues or cells such as podocytes, parietal cells, brush bordercells, glomeruli, nephrons, proximal tubules, distal tubules, collectingducts, interstitial cells, Bowman's capsule, the Loop of Henle, thekidney cortex, the kidney medulla, the calyces, the renal pelvis, kidneyconnective tissue, blood vessels, peripheral nerves, fibroblasts,monocytes/macrophages, lymphocytes, plasma cells, adipocytes,endothelial cells, neurons, or any combination thereof. Thecystine-dense peptide conjugated to a drug can bind to, home to, migrateto, accumulate in, be retained by, or be directed to a kidney and itscomponents. Additionally, in some embodiments, cystine-dense peptidescan penetrate into cells. In other embodiments, cystine-dense peptidesdo not enter cells. In other embodiments, cystine-dense peptides exhibitmore rapid clearance and cellular uptake compared to other types ofmolecules.

The peptides of the present disclosure can comprise cysteine amino acidresidues. In some cases, the peptide has at least 4 cysteine amino acidresidues. In some cases, the peptide has at least 6 cysteine amino acidresidues. In other cases, the peptide has at least 8 cysteine amino acidresidues, at least 10 cysteine amino acid residues, at least 12 cysteineamino acid residues, at least 14 cysteine amino acid residues or atleast 16 cysteine amino acid residues.

A cystine-dense peptide can comprise disulfide bridges. A cystine-densepeptide can be a peptide wherein 5% or more of the residues arecysteines forming intramolecular disulfide bonds as cystines. Adisulfide-linked peptide can be a drug scaffold. In some embodiments,the disulfide bridges form an inhibitor knot. A disulfide bridge can beformed between cysteine residues, for example, between cysteines 1 and4, 2 and 5, or 3 and 6. In some cases, one disulfide bridge passesthrough a loop formed by the other two disulfide bridges, for example,to form the inhibitor knot. In other cases, the disulfide bridges can beformed between any two cysteine residues.

The present disclosure further includes peptide scaffolds that, e.g.,can be used as a starting point for generating additional peptides thatcan target and home to a kidney. In some embodiments, these scaffoldscan be derived from a variety of cystine-dense peptides. In certainembodiments, cystine-dense peptides are assembled into a complextertiary structure that is characterized by a number of intramoleculardisulfide crosslinks, and optionally contain beta strands and othersecondary structures such as an alpha helix. For example, cystine-densepeptides include, in some embodiments, small disulfide-rich proteinscharacterized by a disulfide through disulfide knot. This knot can be,e.g., obtained when one disulfide bridge crosses the macrocycle formedby two other disulfides and the interconnecting backbone. In someembodiments, the cystine-dense peptides can include growth factorcysteine knots or inhibitor cysteine knots. Other possible peptidestructures can include peptide having two parallel helices linked by twodisulfide bridges without β-sheets (e.g., hefutoxin).

A cystine-dense peptide can comprise at least one amino acid residue inan L configuration. A cystine-dense peptide can comprise at least oneamino acid residue in a D configuration. In some embodiments, acystine-dense peptide is 22-63 amino acid residues long. In someembodiments, a cystine-dense peptide is 15-40 amino acid residues long.In other embodiments, a cystine-dense peptide is 11-57 amino acidresidues long. In further embodiments, a cystine-dense peptide is atleast 20 amino acid residues long.

In certain embodiments, the peptides of the present disclosure compriseor are derived from a human protein or peptide that comprises acystine-dense peptide. Examples of such human proteins or peptidesinclude but are not limited to: bone morphogenic protein 7, gremlin,Cerberus, human chorionic gonadotrophin (hCG), AgRP, siderocalin,receptor-associated protein (RAP), ANKRA2, LRP2BP, DAB2, lactoferrin,and other known megalin/cubulin interactors. Optionally, the humanproteins or peptides provided herein are used for motif grafting ontocystine-dense peptide scaffolds.

In alternative embodiments, the peptides of the present disclosurecomprise or are derived from a non-human protein or peptide thatcomprises a cystine-dense peptide, but are modified to include aminoacid sequences found in human proteins or peptides. Such modificationscan be performed in order to enable binding to human targets (e.g.,grafting a known epitope from a human protein that binds to themegalin/cubulin receptor in order to promote proximal tubule binding).

In some embodiments, the peptides of the present disclosure comprise oneor more cysteine amino acid residues. In certain embodiments, thepeptide comprises at least 4, at least 5, at least 6, at least 7, atleast 8, at least 9, at least 10, at least 11, at least 12, at least 13,at least 14, at least 15, or at least 16 cysteine residues.

A cystine-dense peptide can comprise disulfide bridges. A cystine-densepeptide can be a peptide wherein 5% or more of the residues arecysteines forming intramolecular disulfide bonds. A disulfide-linkedpeptide can be a drug scaffold. In some embodiments, the peptides of thepresent disclosure comprise a plurality of disulfide bridges forming aninhibitor knot. In certain embodiments, the disulfide bridges are formedbetween cysteine residues of the peptide. For example, in variousembodiments, the 1^(st) cysteine residue in the sequence is disulfidebonded with the 4^(th) cysteine residue in the sequence, the 2^(nd)cysteine residue in the sequence is disulfide bonded with the 5^(th)cysteine residue in the sequence, and/or the 3^(rd) cysteine residue inthe sequence is disulfide bonded with the 6^(th) cysteine residue in thesequence. In alternative embodiments, the disulfide bridges can beformed between any two cysteine residues. In some cases, one disulfidebridge passes through a loop or ring formed by two other disulfidebridges, for example, to form a disulfide through disulfide knot (e.g.,an inhibitor knot), also known as a “two-and-through” system.

In some embodiments, the peptide contains one or more disulfide bondsand has a positive net charge at neutral pH, where the net charge of thepeptide is greater than or equal to 0 and less than or equal to +30 orwhere the net charge of the peptide is greater than or equal to −30 andless than or equal to 0. For example, in some embodiments, the peptidehas a positive net charge at neutral pH, where the net charge is +0.5 orless than +0.5, +1 or less than +1, +1.5 or less than +1.5, +2 or lessthan +2, +2.5 or less than +2.5, +3 or less than +3, +3.5 or less than+3.5, +4 or less than +4, +4.5 or less than +4.5, +5 or less than +5,+5.5 or less than +5.5, +6 or less than +6, +6.5 or less than +6.5, +7or less than +7, +7.5 or less than +7.5, +8 or less than +8, +8.5 orless than +8.5, +9 or less than +9.5, +10 or less than +10, +11 or lessthan +11, +12 or less than +12, +13 or less than +13, +14 or less than+14, +15 or less than +15, +16 or less than +16, +17 or less than +17,+18 or less than +18, +19 or less than +19, +20 or less than +20, +21 orless than +21, +22 or less than +22, +23 or less than +23, +24 or lessthan +24, +25 or less than +25, +26 or less than +26, +27 or less than+27, +28 or less than +28, +29 or less than +29, or +30 or less than+30. In some embodiments, the peptide has a negative net charge atneutral pH, where the net charge is −0.5 or more than −0.5, −1 or morethan −1, −1.5 or more than −1.5, −2 or more than −2, −2.5 or more than−2.5, −3 or more than −3, −3.5 or more than −3.5, −4 or more than −4,−4.5 or more than −4.5, −5 or more than −5, −5.5 or more than −5.5, −6or more than −6, −6.5 or more than −6.5, −7 or more than −7, −7.5 ormore than −7.5, −8 or more than −8, −8.5 or more than −8.5, −9 or morethan −9.5, −10 or more than −10, −11 or more than −11, −12 or more than−12, −13 or more than −13, −14 or more than −14, −15 or more than −15,−16 or more than −16, −17 or more than −17, −18 or more than −18, −19 ormore than −19, −20 or more than −20, −21 or more than −21, −22 or morethan −22, −23 or more than −23, −24 or more than −24, −25 or more than−25, −26 or more than −26, −27 or more than −27, −28 or more than −28,−29 or more than −29, or −30 or more than −30.

In various embodiments, the peptides of the present disclosure comprisepositively charged amino acid residues. In some embodiments, the peptidehas at least 1 positively charged residue, at least 2 positively chargedresidues, at least 3 positively charged residues, at least 4 positivelycharged residues, at least 5 positively charged residues, at least 6positively charged residues, at least 7 positively charged residues, atleast 8 positively charged residues, at least 9 positively chargedresidues, at least 10 positively charged residues, at least 11positively charged residues, at least 12 positively charged residues, atleast 13 positively charged residues, at least 14 positively chargedresidues, at least 15 positively charged residues, at least 16positively charged residues, or at least 17 positively charged residues.While the positively charged residues can be selected from anypositively charged amino acid residues, in certain embodiments, thepositively charged residues are either K, or R or a combination of K andR.

In various embodiments, the peptides of the present disclosure comprisenegative amino acid residues. In some embodiments, the peptide has 1 orfewer negative amino acid residues, 2 or fewer negative amino acidresidues, 3 or fewer negative amino acid residues, or 4 or fewernegative amino acid residues, 5 or fewer negative amino acid residues, 6or fewer negative amino acid residues, 7 or fewer negative amino acidresidues, 8 or fewer negative amino acid residues, 9 or fewer negativeamino acid residues, or 10 or fewer negative amino acid residues. Whilenegative amino acid residues can be selected from any negative chargedamino acid residues, in certain embodiments, the negative amino acidresidues are either E, or D or a combination of both E and D.

In various embodiments, the peptides of the present disclosure compriseneutral amino acid residues. In some embodiments, the peptide has 1 orfewer neutral amino acid residues, 2 or fewer neutral amino acidresidues, 3 or fewer neutral amino acid residues, 4 or fewer neutralamino acid residues, 5 or fewer neutral amino acid residues, 6 or fewerneutral amino acid residues, 7 or fewer neutral amino acid residues, 8or fewer neutral amino acid residues, 9 or fewer neutral amino acidresidues, 10 or fewer neutral amino acid residues, 15 or fewer neutralamino acid residues, 20 or fewer neutral amino acid residues, 25 orfewer neutral amino acid residues, 30 or fewer neutral amino acidresidues, 35 or fewer neutral amino acid residues, 40 or fewer neutralamino acid residues, or 60 or fewer neutral amino acid residues. In someembodiments, the peptides are members of the pfam00451:toxin_2 family.The pfam00451:toxin_2 structural class family can include a peptide ofany one of IKCSESYQCFPVCKSRFGKTNGRCVNGFCDCF (SEQ ID NO: 577);VKCSSPQQCLKPCKAAFGISAGGKCINGKCKCY (SEQ ID NO: 578);VSCSASSQCWPVCKKLFGTYRGKCMNSKCRCY (SEQ ID NO: 579);ESCTASNQCWSICKRLHNTNRGKCMNKKCRCY (SEQ ID NO: 580);VSCTTSKECWSVCEKLYNTSRGKCMNKKCRCY (SEQ ID NO: 581);MRCKSSKECLVKCKQATGRPNGKCMNRKCKCY (SEQ ID NO: 582);IKCTLSKDCYSPCKKETGCPRAKCINRNCKCY (SEQ ID NO: 583);IRCSGSRDCYSPCMKQTGCPNAKCINKSCKCY (SEQ ID NO: 584);IRCSGTRECYAPCQKLTGCLNAKCMNKACKCY (SEQ ID NO: 585);ISCTNPKQCYPHCKKETGYPNAKCMNRKCKCF (SEQ ID NO: 586);ASCRTPKDCADPCRKETGCPYGKCMNRKCKCN (SEQ ID NO: 587);TSCISPKQCTEPCRAKGCKHGKCMNRKCHCM (SEQ ID NO: 588);KECTGPQHCTNFCRKN-KCTHGKCMNRKCKCF (SEQ ID NO: 589);IKCRTPKDCADPCRKQTGCPHAKCMNKTCRCH (SEQ ID NO: 590);VKCTTSKECWPPCKAATGKAAGKCMNKKCKCQ (SEQ ID NO: 591);LECGASRECYDPCFKAFGRAHGKCMNNKCRCY (SEQ ID NO: 592);EKCFATSQCWTPCKKAIGSLQSKCMNGKCKCY (SEQ ID NO: 593);VRCYASRECWEPCRRVTGSAQAKCQNNQCRCY (SEQ ID NO: 594);VKCSASRECWVACKKVTGSGQGKCQNNQCRCY (SEQ ID NO: 595);VKCISSQECWIACKKVTGRFEGKCQNRQCRCY (SEQ ID NO: 596);VRCYDSRQCWIACKKVTGSTQGKCQNKQCRCY (SEQ ID NO: 597);VDCTVSKECWAPCKAAFGVDRGKCMGKKCKCY (SEQ ID NO: 598);AKCRGSPECLPKCKEAIGKAAGKCMNGKCKCY (SEQ ID NO: 599);KKCQGGSCASVCRRVIGVAAGKCINGRCVCY (SEQ ID NO: 600);KKCSNTSQCYKTCEKVVGVAAGKCMNGKCICY (SEQ ID NO: 601);VKCSGSSKCVKICIDRYNTRGAKCINGRCTCY (SEQ ID NO: 602);NRCNNSSECIPHCIRIFGTRAAKCINRKCYCY (SEQ ID NO: 603);KECNGSSECYSHCEGITGKRSGKCINKKCYCY (SEQ ID NO: 604);AFCNLRRCELSCRSLGLLGKCIGEECKCV (SEQ ID NO: 605);AVCNLKRCQLSCRSLGLLGKCIGDKCECV (SEQ ID NO: 606);AACYSS-DCRVKCVAMGFSSGKCINSKCKCY (SEQ ID NO: 607);AICATDADCSRKCPGNPPCRNGFCACT (SEQ ID NO: 608);TECQIKNDCQRYCQSVKECKYGKCYCN (SEQ ID NO: 609);TQCQSVRDCQQYCLTPDRCSYGTCYCK (SEQ ID NO: 610);VSCRYGSDCAEPCKRLKCLLPSKCINGKCTCY (SEQ ID NO: 611);IKCRYPADCHIMCRKVTGRAEGKCMNGKCTCY (SEQ ID NO: 612);IKCSSSSSCYEPCRGVTGRAHGKCMNGRCTCY (SEQ ID NO: 613);VKCTGSKQCLPACKAAVGKAAGKCMNGKCKCY (SEQ ID NO: 614);VSCKHSGQCIKPCKDA-GMRFGKCMNRKCDCT (SEQ ID NO: 615);VKCRGSPQCIQPCRDA-GMRFGKCMNGKCHCT (SEQ ID NO: 616);VKCTSPKQCLPPCKAQFGIRAGAKCMNGKCKCY (SEQ ID NO: 617);VKCTSPKQCSKPCKELYGSSAGAKCMNGKCKCY (SEQ ID NO: 618);VKCTSPKQCLPPCKEIYGRHAGAKCMNGKCHCS (SEQ ID NO: 619);VKCTGSKQCWPVCKQMFGKPNGKCMNGKCRCY (SEQ ID NO: 620);VKCRGSRDCLDPCKKAGMRFGKCINSKCHCT (SEQ ID NO: 621);VRCVTDDDCFRKCPGNPSCKRGFCACK (SEQ ID NO: 622); orVPCNNSRPCVPVCIREVNNKNGKCSNGKCLCY (SEQ ID NO: 623). A kidney homingpeptide of this disclosure can be a variant of any peptide members ofthe pfam00451:toxin_2 family. In some embodiments, an exemplary kidneyhoming peptide of this disclosure that is a variant of thepfam00451:toxin_2 structural class family is a peptide of SEQ ID NO: 45.In other embodiments, an exemplary kidney homing peptide of thisdisclosure that is a variant of the pfam00451:toxin_2 structural classfamily is a peptide of SEQ ID NO: 132. In other embodiments, the variantpeptides are at least 30% identical to a peptide of the structural classpfam00451:toxin_2 family. In some embodiments, the variant peptides are30%, 40%, 50%, 60%, 80%, 90% or 95% identical to a peptide of thestructural class pfam00451:toxin_2 family. In some embodiments, thevariant peptides are at least 30%, at least 40%, at least 50%, at least60%, at least 80%, at least 90% or at least 95% identical to a peptideof the structural class pfam00451:toxin_2 family.

In some embodiments, kidney homing peptides are family members of thesequences

(SEQ ID NO: 538) GSXVXXXVKCXGSKQCXXPCKRXXGXRXGKCINKKXCKCYXXX or(SEQ ID NO: 558) XVXXXVKCXGSKQCXXPCKRXXGXRXGKCINKKXCKCYXXX,wherein X can be any amino acid, amino acid analogue, or null, in whichthese sequences are based on the most common elements found in thefollowing sequences: GSGVPINVKCRGSRDCLDPCKKA-GMRFGKCINSK-CHCTP-- (SEQ IDNO: 45), GS-VRIPVSCKHSGQCLKPCKDA-GMRFGKCMNGK-CDCTPK- (SEQ ID NO: 44),GSQVQTNVKCQGGS-CASVCRREIGVAAGKCINGK-CVCYRN- (SEQ ID NO: 48),GS-----ISCTGSKQCYDPCKRKTGCPNAKCMNKS-CKCYGCG (SEQ ID NO: 47),GSEV---IRCSGSKQCYGPCKQQTGCTNSKCMNKV-CKCYGCG (SEQ ID NO: 49),GSAVCVYRT------CDKDCKRR-GYRSGKCINNA-CKCYPYG (SEQ ID NO: 46),GS----GIVC---KVCKIICGMQ-GKKVNICKAPIKCKCKKG- (SEQ ID NO: 42), andGSQIYTSKECNGSSECYSHCEGITGKRSGKCINKK-CYCYR-- (SEQ ID NO: 51), where thefollowing residues may be independently interchanged in the sequences: Kand R; M, I, L, and V; G and A; S and T; Q and N; and X canindependently be any number of any amino acid or no amino acid. TheN-terminal GS sequence can be included or excluded between the peptidesof the present disclosure.

In other embodiments, peptides are family members of the sequencesGSXXXGCVXXXXKCRPGXKXCCXPXKRCSRRFGXXXXKKCKXXXXXX (SEQ ID NO: 539) orXXXGCVXXXXKCRPGXKXCCXPXKRCSRRFGXXXXKKCKXXXXXX (SEQ ID NO: 559), in whichthe sequence is based on the most common elements found in the followingsequences: GS---ACKGVFDACTPGKNECC-PNRVCSDK-H----KWCKWKL--- (SEQ ID NO:50), GS---GCLEFWWKCNPNDDKCCRPKLKCSKLF-----KLCNFSFG-- (SEQ ID NO: 52),GSSEKDCIKHLQRCR-ENKDCC--SKKCSRR-GTNPEKRCR------ (SEQ ID NO: 43), andGS---GCFGY--KCDYY-KGCCSGYV-CSPTW-----KWCVRPGPGR (SEQ ID NO: 54), wherethe following residues may be independently interchanged in thesequences: K and R; M, I, L, and V; G and A; S and T; Q and N; and X canindependently be any number of any amino acid or no amino acid. TheN-terminal GS sequence can be included or excluded between the peptidesof the present disclosure.

In some embodiments, a peptide comprises the sequenceGSGVX¹IX²X³KCX⁴GSKQCX⁵DPCKX⁶X⁷X⁸GX⁹RX¹⁰GKCX¹¹NKKCKCX¹²X¹³X¹⁴X¹⁵ (SEQ IDNO: 530) orGVX¹IX²X³KCX⁴GSKQCX⁵DPCKX⁶X⁷X⁸GX⁹RX¹⁰GKCX¹¹NKKCKCX¹²X¹³X¹⁴X¹⁵ (SEQ IDNO: 550), wherein X¹, X², X³, X⁴, X⁵, X⁶, X⁷, X⁸, X⁹, X¹⁰, X¹¹, X¹²,X¹³, X¹⁴ and X¹⁵ are each individually any amino acid or amino acidanalogue or null. In some cases, the peptide comprises the sequenceGSGVX¹IX²X³KCX⁴GSKQCX⁵DPCKX⁶X⁷X⁸GX⁹RX¹⁰GKCX¹¹NKKCKCX¹²X¹³X¹⁴X¹⁵ (SEQ IDNO: 531) orGVX¹IX²X³KCX⁴GSKQCX⁵DPCKX⁶X⁷X⁸GX⁹RX¹⁰GKCX¹¹NKKCKCX¹²X¹³X¹⁴X¹⁵ (SEQ IDNO: 551), where X¹ is selected from P or R, wherein X² is selected fromP or N, wherein X³ is selected from V or I, wherein X⁴ is selected fromS, T, R or K, wherein X⁵ is selected from Y or L, wherein X⁶ is selectedfrom Q, R or K, wherein X⁷ is selected from A, K or R, wherein X⁸ isselected from T or A, wherein X⁹ is selected from C or M, wherein X¹⁰ isselected from F or N, wherein X¹¹ is selected from M or I, wherein X¹²is selected from Y or T, wherein X¹³ is selected from G or P, whereinX¹⁴ is selected from C or null, and wherein X¹⁵ is selected from G ornull.

In some embodiments, a peptide comprises the sequenceGSX¹X²X³X⁴IX⁵CX⁶GSKQCYX⁷PCKX⁸X⁹TGCX¹⁰X¹¹X¹²KCX¹³X¹⁴KX¹⁵CKCYGCG (SEQ IDNO: 532) or X¹X²X³X⁴IX⁵CX⁶GSKQCYX⁷PCKX⁸X⁹TGCX¹⁰X¹¹X¹²KCX¹³X¹⁴KX¹⁵CKCYGCG(SEQ ID NO: 552), wherein X¹, X², X³, X⁴, X⁵, X⁶, X⁷, X⁸, X⁹, X¹⁰, X¹¹,X¹², X¹³, X¹⁴, and X¹⁵ are each individually any amino acid or aminoacid analogue or null. In some cases, the peptide comprises the sequenceGSX¹X²X³X⁴IX⁵CX⁶GSKQCYX⁷PCKX⁸X⁹TGCX¹⁰X¹¹X¹²KCX¹³X¹⁴KX¹⁵CKCYGCG (SEQ IDNO: 533) or X¹X²X³X⁴IX⁵CX⁶GSKQCYX⁷PCKX⁸X⁹X¹⁰X¹¹X¹²KCX¹³X¹⁴KX¹⁵CKCYGCG(SEQ ID NO: 553), where X¹ is selected from G or null, wherein X² isselected from S or null, wherein X³ is selected from E, G or null,wherein X⁴ is selected from V, S, or null, wherein X⁵ is selected from Ror S, wherein X⁶ is selected from S or T, wherein X⁷ is selected from Gor D, wherein X⁸ is selected from Q or R, wherein X⁹ is selected from Qor K, wherein X¹⁰ is selected from T or P, wherein X¹¹ is selected fromN or Q, wherein X¹² is selected from S or A, wherein X¹³ is selectedfrom M or L, wherein X¹⁴ is selected from N or Q, and wherein X¹⁵ isselected from V or S.

In some embodiments, a peptide comprises the sequenceGSX¹X²X³VX⁴IX⁵VX⁶CX⁷X⁸SX⁹X¹⁰CLX¹¹PCKX¹²AGMRFGKCX¹³NX¹⁴KCX¹⁵CTPX¹⁶ (SEQID NO: 534) orX¹X²X³VX⁴IX⁵VX⁶CX⁷X⁸SX⁹X¹⁰CLX¹¹PCKX¹²AGMRFGKCX¹³NX¹⁴KCX¹⁵CTPX¹⁶ (SEQ IDNO: 554)_(, wherein X) ¹, X², X³, X⁴, X⁵, X⁶, X⁷, X⁸, X⁹, X¹⁰, X¹¹, X¹²,X¹³, X¹⁴, X¹⁵, X¹⁶ are each individually any amino acid or amino acidanalogue or null. In some cases, the peptide comprises the sequenceGSX¹X²X³VX⁴IX⁵VX⁶CX⁷X⁸SX⁹X¹⁰CLX¹¹PCKX¹²AGMRFGKCX¹³NX¹⁴KCX¹⁵CTPX¹⁶ (SEQID NO: 535) orX¹X²X³VX⁴IX⁵VX⁶CX⁷X⁸SX⁹X¹⁰CLX¹¹PCKX¹²AGMRFGKCX¹³NX¹⁴KCX¹⁵CTPX¹⁶ (SEQ IDNO: 555), where X¹ is selected from G or null, wherein X² is selectedfrom G, S or null, wherein X³ is selected from G, S or null, wherein X⁴is selected from P or R, wherein X⁵ is selected from N or P, wherein X⁶is selected from K or S, wherein X⁷ is selected from R or K, wherein X⁸is selected from G or H, wherein X⁹ is selected from R or G, wherein X¹⁰is selected from D or Q, wherein X¹¹ is selected from D or K, whereinX¹² is selected from K or D, wherein X¹³ is selected from I or M,wherein X¹⁴ is selected from S or G, wherein X¹⁵ is selected from H orD, and wherein X¹⁶ is selected from K or null.

In some embodiments, a peptide comprises the sequenceGSXVXVKCXGSKQCXPCKRXGXRXGKCINKKXCKCYX (SEQ ID NO: 536) orGSXGCVXKCRPGXKXCCXPXKRCSRRFGXKKCKX (SEQ ID NO: 537), wherein each letteris each individually any amino acid or amino acid analogue and where Xis no amino acid or a 1-10 amino acid long peptide fragment wherein eachamino acid within such peptide fragment can in each case be any aminoacid or amino acid analogue. In some embodiments, a peptide comprisesthe sequence XVXVKCXGSKQCXPCKRXGXRXGKCINKKXCKCYX (SEQ ID NO: 556) orXGCVXKCRPGXKXCCXPXKRCSRRFGXKKCKX (SEQ ID NO: 557), wherein each letteris each individually any amino acid or amino acid analogue and where Xis no amino acid or a 1-10 amino acid long peptide fragment wherein eachamino acid within such peptide fragment can in each case be any aminoacid or amino acid analogue.

In some embodiments, a peptide comprises the sequenceGSGVX¹IX²X³RCX⁴GSRQCX⁵DPCRX⁶X⁷X⁸GX⁹RX¹⁰GRCX¹¹NRRCRCX¹²X¹³X¹⁴X¹⁵ (SEQ IDNO: 540) orGVX¹IX²X³RCX⁴GSRQCX⁵DPCRX⁶X⁷X⁸GX⁹RX¹⁰GRCX¹¹NRRCRCX¹²X¹³X¹⁴X¹⁵ (SEQ IDNO: 560), wherein X¹, X², X³, X⁴, X⁵, X⁶, X⁷, X⁸, X⁹, X¹⁰, X¹¹, X¹²,X¹³, X¹⁴ and X¹⁵ are each individually any amino acid or amino acidanalogue or null. In some cases, the peptide comprises the sequenceGSGVX¹IX²X³RCX⁴GSRQCX⁵DPCRX⁶X⁷X⁸GX⁹RX¹⁰GRCX¹¹NRRCRCX¹²X¹³X¹⁴X¹⁵ (SEQ IDNO: 541) orGVX¹IX²X³RCX⁴GSRQCX⁵DPCRX⁶X⁷X⁸GX⁹RX¹⁰GRCX¹¹NRRCRCX¹²X¹³X¹⁴X¹⁵ (SEQ IDNO: 561), where X¹ is selected from P or R, wherein X² is selected fromP or N, wherein X³ is selected from V or I, wherein X⁴ is selected fromS, T, R or K, wherein X⁵ is selected from Y or L, wherein X⁶ is selectedfrom Q, R or K, wherein X⁷ is selected from A, K or R, wherein X⁸ isselected from T or A, wherein X⁹ is selected from C or M, wherein X¹⁰ isselected from F or N, wherein X¹¹ is selected from M or I, wherein X¹²is selected from Y or T, wherein X¹³ is selected from G or P, whereinX¹⁴ is selected from C or null, and wherein X¹⁵ is selected from G ornull.

In some embodiments, a peptide comprises the sequenceGSX¹X²X³X⁴IX⁵CX⁶GSRQCYX⁷PCRX⁸X⁹TGCX₁₀X¹¹X¹²RCX¹³X¹⁴RX¹⁵CRCYGCG (SEQ IDNO: 542) or X¹X²X³X⁴IX⁵CX⁶GSRQCYX⁷PCRX⁸X⁹TGCX¹⁰X¹¹X¹²RCX¹³X¹⁴RX¹⁵CRCYGCG(SEQ ID NO: 562), wherein X¹, X², X³, X⁴, X⁵, X⁶, X⁷, X⁸, X⁹, X¹⁰, X¹¹,X¹², X¹³, X¹⁴, and X¹⁵ are each individually any amino acid or aminoacid analogue or null. In some cases, the peptide comprises the sequenceGSX¹X²X³X⁴IX⁵CX⁶GSRQCYX⁷PCRX⁸X⁹TGCX¹⁰X¹¹X¹²RCX¹³X¹⁴RX¹⁵CRCYGCG, (SEQ IDNO: 543) or X¹X²X³X⁴IX⁵CX⁶GSRQCYX⁷PCRX⁸X⁹TGCX¹⁰X¹¹X¹²RCX¹³X¹⁴RX¹⁵CRCYGCG(SEQ ID NO: 563), where X¹ is selected from G or null, wherein X² isselected from S or null, wherein X³ is selected from E, G or null,wherein X⁴ is selected from V, S, or null, wherein X⁵ is selected from Ror S, wherein X⁶ is selected from S or T, wherein X⁷ is selected from Gor D, wherein X⁸ is selected from Q or R, wherein X⁹ is selected from Q,R, or K, wherein X¹⁰ is selected from T or P, wherein X¹¹ is selectedfrom N or Q, wherein X¹² is selected from S or A, wherein X¹³ isselected from M or L, wherein X¹⁴ is selected from N or Q, and whereinX¹⁵ is selected from V or S.

In some embodiments, a peptide comprises the sequenceGSX¹X²X³VX⁴IX⁵VX⁶CX⁷X⁸SX⁹X¹⁰CLX¹¹PCRX¹²AGMRFGRCX¹³NX¹⁴RCX¹⁵CTPX¹⁶ (SEQID NO: 544) orX¹X²X³VX⁴IX⁵VX⁶CX⁷X⁸SX⁹X¹⁰CLX¹¹PCRX¹²AGMRFGRCX¹³NX¹⁴RCX¹⁵CTPX¹⁶ (SEQ IDNO: 564), wherein X¹, X², X³, X⁴, X⁵, X⁶, X⁷, X⁸, X⁹, X¹⁰, X¹¹, X¹²,X¹³, X¹⁴, X¹⁵, X¹⁶ are each individually any amino acid or amino acidanalogue or null. In some cases, the peptide comprises the sequenceGSX¹X²X³VX⁴IX⁵VX⁶CX⁷X⁸SX⁹X¹⁰CLX¹¹PCRX¹²AGMRFGRCX¹³NX¹⁴RCX¹⁵CTPX¹⁶ (SEQID NO: 545) orX¹X²X³VX⁴IX⁵VX⁶CX⁷X⁸SX⁹X¹⁰CLX¹¹PCRX¹²AGMRFGRCX¹³NX¹⁴RCX¹⁵CTPX¹⁶ (SEQ IDNO: 565), where X¹ is selected from G or null, wherein X² is selectedfrom G, S or null, wherein X³ is selected from G, S or null, wherein X⁴is selected from P or R, wherein X⁵ is selected from N or P, wherein X⁶is selected from R, K or S, wherein X⁷ is selected from R or K, whereinX⁸ is selected from G or H, wherein X⁹ is selected from R or G, whereinX¹⁰ is selected from D or Q, wherein X¹¹ is selected from D, R, or K,wherein X¹² is selected from K, R, or D, wherein X¹³ is selected from Ior M, wherein X¹⁴ is selected from S or G, wherein X¹⁵ is selected fromH or D, and wherein X¹⁶ is selected from K, R, or null.

In some embodiments, a peptide comprises the sequenceGSXVXVRCXGSRQCXPCRRXGXRXGRCINRRXCRCYX (SEQ ID NO: 546) orGSXGCVXRCRPGXRXCCXPXRRCSRRFGXRRCRX (SEQ ID NO: 547), wherein each letteris each individually any amino acid or amino acid analogue and where Xis no amino acid or a 1-10 amino acid long peptide fragment wherein eachamino acid within such peptide fragment can in each case be any aminoacid or amino acid analogue. In some embodiments, a peptide comprisesthe sequence XVXVRCXGSRQCXPCRRXGXRXGRCINRRXCRCYX (SEQ ID NO: 566) orXGCVXRCRPGXRXCCXPXRRCSRRFGXRRCRX (SEQ ID NO: 567), wherein each letteris each individually any amino acid or amino acid analogue and where Xis no amino acid or a 1-10 amino acid long peptide fragment wherein eachamino acid within such peptide fragment can in each case be any aminoacid or amino acid analogue.

In some embodiments, a peptide comprises the sequenceGSXVXXXVRCXGSRQCXXPCRRXXGXRXGRCINRRXCRCYXXX (SEQ ID NO: 548),XVXXXVRCXGSRQCXXPCRRXXGXRXGRCINRRXCRCYXXX (SEQ ID NO: 568),GSXXXGCVXXXXRCRPGXRXCCXPXRRCSRRFGXXXXRRCRXXXXXX (SEQ ID NO: 549), orXXXGCVXXXXRCRPGXRXCCXPXRRCSRRFGXXXXRRCRXXXXXX (SEQ ID NO: 569) wherein Xis no amino acid or any amino acid analogue.

In some embodiments, a peptide comprises the one or more of thefollowing peptide fragments: GKCINKKCKC (SEQ ID NO: 356); KCIN (SEQ IDNO: 357); KKCK (SEQ ID NO: 358); PCKR (SEQ ID NO: 359); KRCSRR (SEQ IDNO: 360); KQC (SEQ ID NO: 361); GRCINRRCRC (SEQ ID NO: 442); RCIN (SEQID NO: 443); RRCR (SEQ ID NO: 444); PCRR (SEQ ID NO: 445); RRCSRR (SEQID NO: 446); RQC (SEQ ID NO: 447); PCKK (SEQ ID NO: 449), and KKCSKK(SEQ ID NO: 450).

TABLE 1 lists some exemplary peptides according to the presentdisclosure.

TABLE 1 Exemplary Amino Acid Sequences SEQ ID NO Amino Acid SequenceSEQ ID NO: 1 GSDCLPHLRRCRADNDCCGRRCRRRGTNAERRCR SEQ ID NO: 2GSDCKYKFENWGACDGGTGTKVRQGTLKKARYNAQCQETIRVTKPC SEQ ID NO: 3GSDCKYKFENWGACDGGTGTKVRQGTLKKARYNAQCQETIRVTKPC TPKTKAKAKAKKGKGKDSEQ ID NO: 4 GSSCEPGRTFRDRCNTCRCGADGRSAACTLRACPNQ SEQ ID NO: 5GSQFTNVSCTTSRECWSVCQRLHNTSRGRCMNRRCRCYS SEQ ID NO: 6GSMCMPCFTTDHQMARRCDDCCGGRGRGRCYGPQCLCR SEQ ID NO: 7GSISIGIKCSPSIDLCEGQCRIRKYFTGYCSGDTCHCSG SEQ ID NO: 8GSSCAKPRENCNRMNILCCRGECVCPTFGDCFCYGD SEQ ID NO: 9GSNFKVEGACSKPCRKYCIDKGARNGKCINGRCHCYY SEQ ID NO: 10GSDRDSCIDKSRCSKYGYYQECQDCCKKAGHNGGTCMFFKCKCA SEQ ID NO: 11GSQFCGTNGKPCVNGQCCGALRCVVTYHYADGVCLKMNP SEQ ID NO: 12GSRPTDIKCSASYQCFPVCKSRFGKTNGRCVNGLCDCF SEQ ID NO: 13GSNCAGYMRECKEKLCCSGYVCSSRWKWCVLPAPWRR SEQ ID NO: 14GSQFTDVKCTGSKQCWPVCKQMFGKPNGKCMNGKCRCYS SEQ ID NO: 15GSQIDTNVKCSGSSKCVKICIDRYNTRGAKCINGRCTCYP SEQ ID NO: 16GSAEIIRCSGTRECYAPCQKLTGCLNAKCMNKACKCYGCV SEQ ID NO: 17GSSDYCSNDFCFFSCRRDRCARGDCENGKCVCKNCHLN SEQ ID NO: 18GSCIGEGVPCDENDPRCCFGLVCLKPTLHGIWYKSYYCYKK SEQ ID NO: 19GSSCAKPGEMCMRIKCCDGQCGCNRGTGRCFCK SEQ ID NO: 20GSACLAEYQKCEGSTVPCCPGLSCSAGRFRKTKLCTK SEQ ID NO: 21GSVVIGQRCYRSPDCYSACKKLVGKATGKCTNGRCDC SEQ ID NO: 22GSACQFWSCNSSCISRGYRQGYCWGIQYKYCQCQ SEQ ID NO: 23GSRCPPCFTTNPNMEADCRKCCGGRGYCASYQCICPGG SEQ ID NO: 24GSQVSTNKKCSNTSQCYKTCEKVVGVAAGKCMNGKCICYP SEQ ID NO: 25GSECLEIFKACNPSNDQCCKSSKLVCSRKTRWCKYQIG SEQ ID NO: 26GSQDKCKKVYENYPVSKCQLANQCNYDCKLDKHARSGECFYDEKR NLQCICDYCEY SEQ ID NO: 27GSGHACYRNCWREGNDEETCKERC SEQ ID NO: 28GSMCMPCFTTDTQMQERCDRCCGGGGRGRCWGPQCLCI SEQ ID NO: 29GSMCMPCFTTEQRMAIICDDCCGGFGRGRCYGPQCLCR SEQ ID NO: 30GSICIPCFTTDHQIARRCDDCCGGRGRGRCYGPQCICR SEQ ID NO: 31GSRCQLQGFNCVVRSYGLPTIPCCRGLTCRSYFPGSTYGRCQRY SEQ ID NO: 32GSSFGLCRLRRGFCARGRCRFPSIPIGRCSRFVQCCRRVW SEQ ID NO: 33GSSCEPGTTFRDRCNTCRCGSDGRSAACTLRACPQ SEQ ID NO: 34GSSCTPGTTFRDRCNTCRCSSNGRSAACTLRACPPGSY SEQ ID NO: 35GSSCTPGTTFRNRCNTCRCGSNGRSASCTLMACPPGSY SEQ ID NO: 36GSSCTPGATFRNRCNTCRCGSNGRSASCTLMACPPGSY SEQ ID NO: 37GSSCQPGTTYQRGCNTCRCLEDGQTEACTLRLC SEQ ID NO: 38GSSCTPGATYREGCNICRCRSDGRSGACTRRICPVDSN SEQ ID NO: 39GSSCQPGTTFRRDCNTCVCNRDGTNAACTLRACL SEQ ID NO: 40GGYSRCQLQGFNCVVRSYGLPTIPCCRGLTCRSYFPGSTYGRCQRY SEQ ID NO: 41GSSCARPRENCNRMNILCCRGECVCPTFGDCFCYGD SEQ ID NO: 42GSGIVCKVCKIICGMQGKKVNICKAPIKCKCKKG SEQ ID NO: 43GSSEKDCIKHLQRCRENKDCCSKKCSRRGTNPEKRCR SEQ ID NO: 44GSVRIPVSCKHSGQCLKPCKDAGMRFGKCMNGKCDCTPK SEQ ID NO: 45GSGVPINVKCRGSRDCLDPCKKAGMRFGKCINSKCHCTP SEQ ID NO: 46GSAVCVYRTCDKDCKRRGYRSGKCINNACKCYPYG SEQ ID NO: 47GSISCTGSKQCYDPCKRKTGCPNAKCMNKSCKCYGCG SEQ ID NO: 48GSQVQTNVKCQGGSCASVCRREIGVAAGKCINGKCVCYRN SEQ ID NO: 49GSEVIRCSGSKQCYGPCKQQTGCTNSKCMNKVCKCYGCG SEQ ID NO: 50GSACKGVFDACTPGKNECCPNRVCSDKHKWCKWKL SEQ ID NO: 51GSQIYTSKECNGSSECYSHCEGITGKRSGKCINKKCYCYR SEQ ID NO: 52GSGCLEFWWKCNPNDDKCCRPKLKCSKLFKLCNFSFG SEQ ID NO: 53GSDCVRFWGKCSQTSDCCPHLACKSKWPRNICVWDGSVG SEQ ID NO: 54GSGCFGYKCDYYKGCCSGYVCSPTWKWCVRPGPGR SEQ ID NO: 55GSMNAKFILLLVLTTMMLLPDTKGAEVIRCSGSKQCYGPCKQQTGCT NSKCMNKVCKCYGCGSEQ ID NO: 56 GSMNAKLIYLLLVVTTMTLMFDTAQAVDIMCSGPKQCYGPCKKETGCPNAKCMNRRCKCYGCV SEQ ID NO: 57GSMNAKLIYLLLVVTTMMLTFDTTQAGDIKCSGTRQCWGPCKKQTT CTNSKCMNGKCKCYGCVGSEQ ID NO: 58 GSMNTKFIFLLLVVTNTMMLFDTKPVEGISCTGSKQCYDPCKRKTGCPNAKCMNKSCKCYGCG SEQ ID NO: 59 GSGVPINVKCSGSRDCLEPCKKAGMRFGKCINRKCHCTPKSEQ ID NO: 60 GSGVPINVKCTGSPQCLKPCKDAGMRFGKCINGKCHCTPK SEQ ID NO: 61GSGVIINVKCKISRQCLEPCKKAGMRFGKCMNGKCHCTPK SEQ ID NO: 62GSGVPINVKCRGSPQCIQPCRDAGMRFGKCMNGKCHCTPQ SEQ ID NO: 63GSGVEINVKCTGSHQCIKPCKDAGMRFGKCINRKCHCTPK SEQ ID NO: 64GSGVEINVKCSGSPQCLKPCKDAGMRFGKCMNRKCHCTPK SEQ ID NO: 65GSGVPTDVKCRGSPQCIQPCKDAGMRFGKCMNGKCHCTPK SEQ ID NO: 66GSGVPINVSCTGSPQCIKPCKDAGMRFGKCMNRKCHCTPK SEQ ID NO: 67GSGVPINVPCTGSPQCIKPCKDAGMRFGKCMNRKCHCTPK SEQ ID NO: 68GSVGINVKCKHSGQCLKPCKDAGMRFGKCINGKCDCTPK SEQ ID NO: 69GSVGINVKCKHSGQCLKPCKDAGMRFGKCMNGKCDCTPK SEQ ID NO: 70GSVGIPVSCKHSGQCIKPCKDAGMRFGKCMNRKCDCTPK SEQ ID NO: 71GSRKGCFKEGHSCPKTAPCCRPLVCKGPSPNTKKCTRP SEQ ID NO: 72GSSFCIPFKPCKSDENCCKKFKCKTTGIVKLCRW SEQ ID NO: 73GSLKGCLPRNRFCNALSGPRCCSGLRCKELSIWASKCL SEQ ID NO: 74GSGNYCLRGRCLPGGRKCCNGRPCECFAKICSCKPK SEQ ID NO: 75GSTVKCGGCNRKCCPGGCRSGKCINGKCQCY SEQ ID NO: 76GSGCMKEYCAGQCRGKVSQDYCLKHCKCIPR SEQ ID NO: 77GSACLGFGEKCNPSNDKCCKSSSLVCSQKHKWCKYG SEQ ID NO: 78GSRGGCLPHNRFCNALSGPRCCSGLRCKELSIRDSRCLG SEQ ID NO: 79GSRGGCLPRNKFCNPSSGPRCCSGLTCKELNIWASKCL SEQ ID NO: 80GSQRSCAKPGDMCMGIKCCDGQCGCNRGTGRCFCK SEQ ID NO: 81GSARGCADAYKSCNHPRTCCDGYNGYKRACICSGSNCKCKKS SEQ ID NO: 82GSRGGCLPHNRFCNALSGPRCCSGLRCKELSIWDSRCLG SEQ ID NO: 83GSRGGCLPHNRFCNALSGPRCCSGLKCKELSIYDSRCLG SEQ ID NO: 84GSRGGCLPHNRFCNALSGPRCCSRLKCKELSIWDSRCLG SEQ ID NO: 85GSRGGCLPHNRFCNALTGPRCCSRLRCKELSIWDSICLG SEQ ID NO: 86GSSCADAYKSCDSLKCCNNRTCMCSMIGTNCTCRKK SEQ ID NO: 87GSERRCLPAGKTCVRGPMRVPCCGSCSQNKCT SEQ ID NO: 88GSLCSREGEFCYKLRKCCAGFYCKAFVLHCYRN SEQ ID NO: 89GSACGSCRKKCKGSGKCINGRCKCY SEQ ID NO: 90 GSACGSCRKKCKGPGKCINGRCKCYSEQ ID NO: 91 GSACQGYMRKCGRDKPPCCKKLECSKTWRWCVWN SEQ ID NO: 92GSGRYCQKWMWTCDSKRACCEGLRCKLWCRKI SEQ ID NO: 93GSNAKCRGSPECLPKCKEAIGKAAGKCMNGKCKCYP SEQ ID NO: 94GSNVKCRGSKECLPACKAAVGKAAGKCMNGKCKCYP SEQ ID NO: 95GSNVKCRGSPECLPKCKEAIGKSAGKCMNGKCKCYP SEQ ID NO: 96GSNAKCRGSPECLPKCKQAIGKAAGKCMNGKCKCYP SEQ ID NO: 97GSRGYCAEKGIKCHNIHCCSGLTCKCKGSSCVCRK SEQ ID NO: 98GSERGCKLTFWKCKNKKECCGWNACALGICNIPR SEQ ID NO: 99GSKKKCIAKDYGRCKWGGTPCCRGRGCICSIMGTNCECKPR SEQ ID NO: 100GSGCKLTFWKCKNKKECCGWNACALGICMPR SEQ ID NO: 101GSACKGLFVTCTPGKDECCPNHVCSSKHKWCKYK SEQ ID NO: 102GSIACAPRGLLCFRDKECCKGLTCKGRFVNTWPTFCLV SEQ ID NO: 103GSACAGLYKKCGKGVNTCCENRPCKCDLAMGNCICKKK SEQ ID NO: 104GSFTCAISCDIKVNGKPCKGSGEKKCSGGWSCKFNVCVKV SEQ ID NO: 105GSGFCAQKGIKCHDIECCTNLKCVREGSNRVCRKA SEQ ID NO: 106GSCAKKRNWCGKNEDCCCPMKCIYAWYNQQGSCQSTITGLFKKC SEQ ID NO: 107GSYCQKWMWTCDSARKCCEGLVCRLWCKKI SEQ ID NO: 108GSRGGCLPHNKFCNALSGPRCCSGLKCKELTIWNTKCLE SEQ ID NO: 109GSNVKCTGSKQCLPACKAAVGKAAGKCMNGKCKCYT SEQ ID NO: 110GSQRSCAKPGEMCMRIKCCDGQCGCNRGTGRCFCK SEQ ID NO: 111GSGCIPKHKRCTWSGPKCCNNISCHCNISGTLCKCRPG SEQ ID NO: 112GSNYCVAKRCRPGGRQCCSGKPCACVGKVCKCPRD SEQ ID NO: 113GSERGCSGAYKRCSSSQRCCEGRPCVCSAINSNCKCRKT SEQ ID NO: 114GSRYCPRNPEACYNYCLRTGRPGGYCGGRSRITCFCFR SEQ ID NO: 115GSQRSCAKPGEMCMGIKCCDGQCGCNRGTGRCFCK SEQ ID NO: 116GSRRGCFKEGKWCPKSAPCCAPLKCKGPSIKQQKCVRE SEQ ID NO: 117GSTVKCGGCNRKCCAGGCRSGKCINGKCQCYGR SEQ ID NO: 118GSERRCEPSGKPCRPLMRIPCCGSCVRGKCA SEQ ID NO: 119GSRGGCLPRNKFCNPSSGPRCCSGLTCKELNIWANKCL SEQ ID NO: 120GSCAKKRNWCGKNEDCCCPMKCIYAWYNQQGSCQTTITGLFKKC SEQ ID NO: 127GSVRIPVSCKHSGQCLKPCKDAGMRTGKCMNGKCDCTPK SEQ ID NO: 128GSVKCTTSKDCWPPCKKVTGRA SEQ ID NO: 129 GSGIVCRVCRIICGMQGRRVNICRAPIRCRCRRGSEQ ID NO: 130 GSSERDCIRHLQRCRENRDCCSRRCSRRGTNPERRCR SEQ ID NO: 131GSVRIPVSCRHSGQCLRPCRDAGMRFGRCMNGRCDCTPR SEQ ID NO: 132GSGVPINVRCRGSRDCLDPCRRAGMRFGRCINSRCHCTP SEQ ID NO: 133GSAVCVYRTCDRDCRRRGYRSGRCINNACRCYPYG SEQ ID NO: 134GSISCTGSRQCYDPCRRRTGCPNARCMNRSCRCYGCG SEQ ID NO: 135GSQVQTNVRCQGGSCASVCRREIGVAAGRCINGRCVCYRN SEQ ID NO: 136GSEVIRCSGSRQCYGPCRQQTGCTNSRCMNRVCRCYGCG SEQ ID NO: 137GSACRGVFDACTPGRNECCPNRVCSDRHRWCRWRL SEQ ID NO: 138GSQIYTSRECNGSSECYSHCEGITGRRSGRCINRRCYCYR SEQ ID NO: 139GSGCLEFWWRCNPNDDRCCRPRLRCSRLFRLCNFSFG SEQ ID NO: 140GSDCVRFWGRCSQTSDCCPHLACRSRWPRNICVWDGSVG SEQ ID NO: 141GSGCFGYRCDYYRGCCSGYVCSPTWRWCVRPGPGR SEQ ID NO: 142GSMNARFILLLVLTTMMLLPDTRGAEVIRCSGSRQCYGPCRQQTGCT NSRCMNRVCRCYGCGSEQ ID NO: 143 GSMNARLIYLLLVVTTMTLMFDTAQAVDEVICSGPRQCYGPCRRETGCPNARCMNRRCRCYGCV SEQ ID NO: 144GSMNARLIYLLLVVTTMMLTFDTTQAGDIRCSGTRQCWGPCRRQTTC TNSRCMNGRCRCYGCVGSEQ ID NO: 145 GSMNTRFIFLLLVVTNTMMLFDTRPVEGISCTGSRQCYDPCRRRTGCPNARCMNRSCRCYGCG SEQ ID NO: 146 GSGVPINVRCSGSRDCLEPCRRAGMRFGRCINRRCHCTPRSEQ ID NO: 147 GSGVPINVRCTGSPQCLRPCRDAGMRFGRCINGRCHCTPR SEQ ID NO: 148GSGVIINVRCRISRQCLEPCRRAGMRFGRCMNGRCHCTPR SEQ ID NO: 149GSGVPINVRCRGSPQCIQPCRDAGMRFGRCMNGRCHCTPQ SEQ ID NO: 150GSGVEINVRCTGSHQCIRPCRDAGMRFGRCINRRCHCTPR SEQ ID NO: 151GSGVEINVRCSGSPQCLRPCRDAGMRFGRCMNRRCHCTPR SEQ ID NO: 152GSGVPTDVRCRGSPQCIQPCRDAGMRFGRCMNGRCHCTPR SEQ ID NO: 153GSGVPINVSCTGSPQCIRPCRDAGMRFGRCMNRRCHCTPR SEQ ID NO: 154GSGVPINVPCTGSPQCIRPCRDAGMRFGRCMNRRCHCTPR SEQ ID NO: 155GSVGINVRCRHSGQCLRPCRDAGMRFGRCINGRCDCTPR SEQ ID NO: 156GSVGINVRCRHSGQCLRPCRDAGMRFGRCMNGRCDCTPR SEQ ID NO: 157GSVGIPVSCRHSGQCIRPCRDAGMRFGRCMNRRCDCTPR SEQ ID NO: 158GSRRGCFREGHSCPRTAPCCRPLVCRGPSPNTRRCTRP SEQ ID NO: 159GSSFCIPFRPCRSDENCCRRFRCRTTGIVRLCRW SEQ ID NO: 160GSLRGCLPRNRFCNALSGPRCCSGLRCRELSIWASRCL SEQ ID NO: 161GSGNYCLRGRCLPGGRRCCNGRPCECFARICSCRPR SEQ ID NO: 162GSTVRCGGCNRRCCPGGCRSGRCINGRCQCY SEQ ID NO: 163GSGCMREYCAGQCRGRVSQDYCLRHCRCIPR SEQ ID NO: 164GSACLGFGERCNPSNDRCCRSSSLVCSQRHRWCRYG SEQ ID NO: 165GSRGGCLPHNRFCNALSGPRCCSGLRCRELSIRDSRCLG SEQ ID NO: 166GSRGGCLPRNRFCNPSSGPRCCSGLTCRELNIWASRCL SEQ ID NO: 167GSQRSCARPGDMCMGIRCCDGQCGCNRGTGRCFCR SEQ ID NO: 168GSARGCADAYRSCNHPRTCCDGYNGYRRACICSGSNCRCRRS SEQ ID NO: 169GSRGGCLPHNRFCNALSGPRCCSGLRCRELSIWDSRCLG SEQ ID NO: 170GSRGGCLPHNRFCNALSGPRCCSGLRCRELSIYDSRCLG SEQ ID NO: 171GSRGGCLPHNRFCNALSGPRCCSRLRCRELSIWDSRCLG SEQ ID NO: 172GSRGGCLPHNRFCNALTGPRCCSRLRCRELSIWDSICLG SEQ ID NO: 173GSSCADAYKSCDSLRCCNNRTCMCSMIGTNCTCRRR SEQ ID NO: 174GSERRCLPAGRTCVRGPMRVPCCGSCSQNRCT SEQ ID NO: 175GSLCSREGEFCYRLRRCCAGFYCRAFVLHCYRN SEQ ID NO: 176GSACGSCRRRCRGSGRCINGRCRCY SEQ ID NO: 177 GSACGSCRRRCRGPGRCINGRCRCYSEQ ID NO: 178 GSACQGYMRRCGRDRPPCCRRLECSRTWRWCVWN SEQ ID NO: 179GSGRYCQRWMWTCDSRRACCEGLRCRLWCRRI SEQ ID NO: 180GSNARCRGSPECLPRCREAIGRAAGRCMNGRCRCYP SEQ ID NO: 181GSNVRCRGSRECLPACRAAVGRAAGRCMNGRCRCYP SEQ ID NO: 182GSNVRCRGSPECLPRCREAIGRSAGRCMNGRCRCYP SEQ ID NO: 183GSNARCRGSPECLPRCRQAIGRAAGRCMNGRCRCYP SEQ ID NO: 184GSRGYCAERGIRCHNIHCCSGLTCRCRGSSCVCRR SEQ ID NO: 185GSERGCRLTFWRCRNRRECCGWNACALGICMPR SEQ ID NO: 186GSRRRCIARDYGRCRWGGTPCCRGRGCICSIMGTNCECRPR SEQ ID NO: 187GSGCRLTFWRCRNRRECCGWNACALGICNIPR SEQ ID NO: 188GSACRGLFVTCTPGRDECCPNHVCSSRHRWCRYR SEQ ID NO: 189GSIACAPRGLLCFRDRECCRGLTCRGRFVNTWPTFCLV SEQ ID NO: 190GSACAGLYRRCGRGVNTCCENRPCRCDLAMGNCICRRR SEQ ID NO: 191GSFTCAISCDIRVNGRPCRGSGERRCSGGWSCRFNVCVRV SEQ ID NO: 192GSGFCAQRGIRCHDIHCCTNLRCVREGSNRVCRRA SEQ ID NO: 193GSCARRRNWCGRNEDCCCPMRCIYAWYNQQGSCQSTITGLFRRC SEQ ID NO: 194GSYCQRWMWTCDSARRCCEGLVCRLWCRRI SEQ ID NO: 195GSRGGCLPHNRFCNALSGPRCCSGLRCRELTIWNTRCLE SEQ ID NO: 196GSNVRCTGSRQCLPACRAAVGRAAGRCMNGRCRCYT SEQ ID NO: 197GSQRSCARPGEMCMRIRCCDGQCGCNRGTGRCFCR SEQ ID NO: 198GSGCIPRHRRCTWSGPRCCNNISCHCNISGTLCRCRPG SEQ ID NO: 199GSNYCVARRCRPGGRQCCSGRPCACVGRVCRCPRD SEQ ID NO: 200GSERGCSGAYRRCSSSQRCCEGRPCVCSAINSNCRCRRT SEQ ID NO: 201GSQRSCARPGEMCMGIRCCDGQCGCNRGTGRCFCR SEQ ID NO: 202GSRRGCFREGRWCPRSAPCCAPLRCRGPSIRQQRCVRE SEQ ID NO: 203GSTVRCGGCNRRCCAGGCRSGRCINGRCQCYGR SEQ ID NO: 204GSERRCEPSGRPCRPLMRIPCCGSCVRGRCA SEQ ID NO: 205GSRGGCLPRNRFCNPSSGPRCCSGLTCRELNIWANRCL SEQ ID NO: 206GSCARRRNWCGRNEDCCCPMRCIYAWYNQQGSCQTTITGLFRRC SEQ ID NO: 213GSVRIPVSCRHSGQCLRPCRDAGMRTGRCMNGRCDCTPR SEQ ID NO: 216GSQKILSNRCNNSSECIPHCIRIFGTRAAKCINRKCYCYP SEQ ID NO: 217GSAVCNLKRCQLSCRSLGLLGKCIGDKCECVKHG SEQ ID NO: 218GSISIGIRCSPSIDLCEGQCRIRRYFTGYCSGDTCHCSG SEQ ID NO: 219GSGDCLPHLRRCRENNDCCSRRCRRRGANPERRCR SEQ ID NO: 220GSSCEPGRTFRDRCNTCKCGADGRSAACTLRACPNQ SEQ ID NO: 221GSGDCLPHLKRCKADNDCCGKKCKRRGTNAEKRCR SEQ ID NO: 222GSGDCLPHLKRCKENNDCCSKKCKRRGTNPEKRCR SEQ ID NO: 223GSKDCLKKLKLCKENKDCCSKSCKRRGTNIEKRCR SEQ ID NO: 224GSGDCLPHLKRCKENNDCCSKKCKRRGANPEKRCR SEQ ID NO: 225GSVFINVKCRGSPECLPKCKEAIGKSAGKCMNGKCKCYP SEQ ID NO: 226GSVFINAKCRGSPECLPKCKEAIGKAAGKCMNGKCKCYP SEQ ID NO: 227GSVIINVKCKISRQCLEPCKKAGMRFGKCMNGKCHCTP SEQ ID NO: 228GSVPTDVKCRGSPQCIQPCKDAGMRFGKCMNGKCHCTP SEQ ID NO: 229GSVRIPVSCKHSGQCLKPCKDAGMRFGKCMNGKCDCTP SEQ ID NO: 230GSVRIPVSCRHSGQCLRPCRDAGMRFGRCMNGRCDCTP SEQ ID NO: 231GSTNVSCTTSKECWSVCQRLHNTSRGKCMNKKCRC SEQ ID NO: 232GSNVKCTGSKQCLPACKAAVGKAAGKCMNGKCKC SEQ ID NO: 233GSGVPINVRCRGSRDCLDPCRGAGERHGRCGNSRCHCTP SEQ ID NO: 234GSVRIPVSCRHSGQCLRPCRDAGERHGRCGGGRCDCTPR SEQ ID NO: 235GSQVQTNVRCQGGSCGSVCRREGGGAGGGCGNGRCGCYRN SEQ ID NO: 236DCLPHLRRCRADNDCCGRRCRRRGTNAERRCR SEQ ID NO: 237DCKYKFENWGACDGGTGTKVRQGTLKKARYNAQCQETIRVTKPC SEQ ID NO: 238DCKYKFENWGACDGGTGTKVRQGTLKKARYNAQCQETIRVTKPCTP KTKAKAKAKKGKGKDSEQ ID NO: 239 SCEPGRTFRDRCNTCRCGADGRSAACTLRACPNQ SEQ ID NO: 240QFTNVSCTTSRECWSVCQRLHNTSRGRCMNRRCRCYS SEQ ID NO: 241MCNIPCFTTDHQMARRCDDCCGGRGRGRCYGPQCLCR SEQ ID NO: 242ISIGIKCSPSIDLCEGQCRIRKYFTGYCSGDTCHCSG SEQ ID NO: 243SCAKPRENCNRMNILCCRGECVCPTFGDCFCYGD SEQ ID NO: 244NFKVEGACSKPCRKYCIDKGARNGKCINGRCHCYY SEQ ID NO: 245DRDSCIDKSRCSKYGYYQECQDCCKKAGHNGGTCMFFKCKCA SEQ ID NO: 246QFCGTNGKPCVNGQCCGALRCVVTYHYADGVCLKMNP SEQ ID NO: 247RPTDIKCSASYQCFPVCKSRFGKTNGRCVNGLCDCF SEQ ID NO: 248NCAGYMRECKEKLCCSGYVCSSRWKWCVLPAPWRR SEQ ID NO: 249QFTDVKCTGSKQCWPVCKQMFGKPNGKCMNGKCRCYS SEQ ID NO: 250QIDTNVKCSGSSKCVKICIDRYNTRGAKCINGRCTCYP SEQ ID NO: 251AEIIRCSGTRECYAPCQKLTGCLNAKCMNKACKCYGCV SEQ ID NO: 252SDYCSNDFCFFSCRRDRCARGDCENGKCVCKNCHLN SEQ ID NO: 253CIGEGVPCDENDPRCCFGLVCLKPTLHGIWYKSYYCYKK SEQ ID NO: 254SCAKPGEMCMRIKCCDGQCGCNRGTGRCFCK SEQ ID NO: 255ACLAEYQKCEGSTVPCCPGLSCSAGRFRKTKLCTK SEQ ID NO: 256VVIGQRCYRSPDCYSACKKLVGKATGKCTNGRCDC SEQ ID NO: 257ACQFWSCNSSCISRGYRQGYCWGIQYKYCQCQ SEQ ID NO: 258RCPPCFTTNPNMEADCRKCCGGRGYCASYQCICPGG SEQ ID NO: 259QVSTNKKCSNTSQCYKTCEKVVGVAAGKCMNGKCICYP SEQ ID NO: 260ECLEIFKACNPSNDQCCKSSKLVCSRKTRWCKYQIG SEQ ID NO: 261QDKCKKVYENYPVSKCQLANQCNYDCKLDKHARSGECFYDEKRNL QCICDYCEY SEQ ID NO: 262GHACYRNCWREGNDEETCKERC SEQ ID NO: 263MCMPCFTTDTQMQERCDRCCGGGGRGRCWGPQCLCI SEQ ID NO: 264MCMPCFTTEQRMAIICDDCCGGFGRGRCYGPQCLCR SEQ ID NO: 265ICIPCFTTDHQIARRCDDCCGGRGRGRCYGPQCICR SEQ ID NO: 266RCQLQGFNCVVRSYGLPTIPCCRGLTCRSYFPGSTYGRCQRY SEQ ID NO: 267SFGLCRLRRGFCARGRCRFPSIPIGRCSRFVQCCRRVW SEQ ID NO: 268SCEPGTTFRDRCNTCRCGSDGRSAACTLRACPQ SEQ ID NO: 269SCTPGTTFRDRCNTCRCSSNGRSAACTLRACPPGSY SEQ ID NO: 270SCTPGTTFRNRCNTCRCGSNGRSASCTLMACPPGSY SEQ ID NO: 271SCTPGATFRNRCNTCRCGSNGRSASCTLMACPPGSY SEQ ID NO: 272SCQPGTTYQRGCNTCRCLEDGQTEACTLRLC SEQ ID NO: 273SCTPGATYREGCNICRCRSDGRSGACTRRICPVDSN SEQ ID NO: 274SCQPGTTFRRDCNTCVCNRDGTNAACTLRACL SEQ ID NO: 275YSRCQLQGFNCVVRSYGLPTIPCCRGLTCRSYFPGSTYGRCQRY SEQ ID NO: 276SCARPRENCNRMNILCCRGECVCPTFGDCFCYGD SEQ ID NO: 277GIVCKVCKIICGMQGKKVNICKAPIKCKCKKG SEQ ID NO: 278SEKDCIKHLQRCRENKDCCSKKCSRRGTNPEKRCR SEQ ID NO: 279VRIPVSCKHSGQCLKPCKDAGMRFGKCMNGKCDCTPK SEQ ID NO: 280GVPINVKCRGSRDCLDPCKKAGMRFGKCINSKCHCTP SEQ ID NO: 281AVCVYRTCDKDCKRRGYRSGKCINNACKCYPYG SEQ ID NO: 282ISCTGSKQCYDPCKRKTGCPNAKCMNKSCKCYGCG SEQ ID NO: 283QVQTNVKCQGGSCASVCRREIGVAAGKCINGKCVCYRN SEQ ID NO: 284EVIRCSGSKQCYGPCKQQTGCTNSKCMNKVCKCYGCG SEQ ID NO: 285ACKGVFDACTPGKNECCPNRVCSDKHKWCKWKL SEQ ID NO: 286QIYTSKECNGSSECYSHCEGITGKRSGKCINKKCYCYR SEQ ID NO: 287GCLEFWWKCNPNDDKCCRPKLKCSKLFKLCNFSFG SEQ ID NO: 288DCVRFWGKCSQTSDCCPHLACKSKWPRNICVWDGSVG SEQ ID NO: 289GCFGYKCDYYKGCCSGYVCSPTWKWCVRPGPGR SEQ ID NO: 290MNAKFILLLVLTTMMLLPDTKGAEVIRCSGSKQCYGPCKQQTGCTNS KCMNKVCKCYGCGSEQ ID NO: 291 MNAKLIYLLLVVTTMTLMFDTAQAVDIMCSGPKQCYGPCKKETGCPNAKCMNRRCKCYGCV SEQ ID NO: 292MNAKLIYLLLVVTTMMLTFDTTQAGDIKCSGTRQCWGPCKKQTTCT NSKCMNGKCKCYGCVGSEQ ID NO: 293 MNTKFIFLLLVVTNTMMLFDTKPVEGISCTGSKQCYDPCKRKTGCPNAKCMNKSCKCYGCG SEQ ID NO: 294 GVPINVKCSGSRDCLEPCKKAGMRFGKCINRKCHCTPKSEQ ID NO: 295 GVPINVKCTGSPQCLKPCKDAGMRFGKCINGKCHCTPK SEQ ID NO: 296GVIINVKCKISRQCLEPCKKAGMRFGKCMNGKCHCTPK SEQ ID NO: 297GVPINVKCRGSPQCIQPCRDAGMRFGKCMNGKCHCTPQ SEQ ID NO: 298GVEINVKCTGSHQCIKPCKDAGMRFGKCINRKCHCTPK SEQ ID NO: 299GVEINVKCSGSPQCLKPCKDAGMRFGKCMNRKCHCTPK SEQ ID NO: 300GVPTDVKCRGSPQCIQPCKDAGMRFGKCMNGKCHCTPK SEQ ID NO: 301GVPINVSCTGSPQCIKPCKDAGMRFGKCMNRKCHCTPK SEQ ID NO: 302GVPINVPCTGSPQCIKPCKDAGMRFGKCMNRKCHCTPK SEQ ID NO: 303VGINVKCKHSGQCLKPCKDAGMRFGKCINGKCDCTPK SEQ ID NO: 304VGINVKCKHSGQCLKPCKDAGMRFGKCMNGKCDCTPK SEQ ID NO: 305VGIPVSCKHSGQCIKPCKDAGMRFGKCMNRKCDCTPK SEQ ID NO: 306RKGCFKEGHSCPKTAPCCRPLVCKGPSPNTKKCTRP SEQ ID NO: 307SFCIPFKPCKSDENCCKKFKCKTTGIVKLCRW SEQ ID NO: 308LKGCLPRNRFCNALSGPRCCSGLRCKELSIWASKCL SEQ ID NO: 309GNYCLRGRCLPGGRKCCNGRPCECFAKICSCKPK SEQ ID NO: 310TVKCGGCNRKCCPGGCRSGKCINGKCQCY SEQ ID NO: 311GCMKEYCAGQCRGKVSQDYCLKHCKCIPR SEQ ID NO: 312ACLGFGEKCNPSNDKCCKSSSLVCSQKHKWCKYG SEQ ID NO: 313RGGCLPHNRFCNALSGPRCCSGLRCKELSIRDSRCLG SEQ ID NO: 314RGGCLPRNKFCNPSSGPRCCSGLTCKELNIWASKCL SEQ ID NO: 315QRSCAKPGDMCMGIKCCDGQCGCNRGTGRCFCK SEQ ID NO: 316ARGCADAYKSCNHPRTCCDGYNGYKRACICSGSNCKCKKS SEQ ID NO: 317RGGCLPHNRFCNALSGPRCCSGLRCKELSIWDSRCLG SEQ ID NO: 318RGGCLPHNRFCNALSGPRCCSGLKCKELSIYDSRCLG SEQ ID NO: 319RGGCLPHNRFCNALSGPRCCSRLKCKELSIWDSRCLG SEQ ID NO: 320RGGCLPHNRFCNALTGPRCCSRLRCKELSIWDSICLG SEQ ID NO: 321SCADAYKSCDSLKCCNNRTCMCSMIGTNCTCRKK SEQ ID NO: 322ERRCLPAGKTCVRGPMRVPCCGSCSQNKCT SEQ ID NO: 323LCSREGEFCYKLRKCCAGFYCKAFVLHCYRN SEQ ID NO: 324 ACGSCRKKCKGSGKCINGRCKCYSEQ ID NO: 325 ACGSCRKKCKGPGKCINGRCKCY SEQ ID NO: 326ACQGYMRKCGRDKPPCCKKLECSKTWRWCVWN SEQ ID NO: 327GRYCQKWMWTCDSKRACCEGLRCKLWCRKI SEQ ID NO: 328NAKCRGSPECLPKCKEAIGKAAGKCMNGKCKCYP SEQ ID NO: 329NVKCRGSKECLPACKAAVGKAAGKCMNGKCKCYP SEQ ID NO: 330NVKCRGSPECLPKCKEAIGKSAGKCMNGKCKCYP SEQ ID NO: 331NAKCRGSPECLPKCKQAIGKAAGKCMNGKCKCYP SEQ ID NO: 332RGYCAEKGIKCHNIHCCSGLTCKCKGSSCVCRK SEQ ID NO: 333ERGCKLTFWKCKNKKECCGWNACALGICMPR SEQ ID NO: 334KKKCIAKDYGRCKWGGTPCCRGRGCICSIMGTNCECKPR SEQ ID NO: 335GCKLTFWKCKNKKECCGWNACALGICMPR SEQ ID NO: 336ACKGLFVTCTPGKDECCPNHVCSSKHKWCKYK SEQ ID NO: 337IACAPRGLLCFRDKECCKGLTCKGRFVNTWPTFCLV SEQ ID NO: 338ACAGLYKKCGKGVNTCCENRPCKCDLAMGNCICKKK SEQ ID NO: 339FTCAISCDIKVNGKPCKGSGEKKCSGGWSCKFNVCVKV SEQ ID NO: 340GFCAQKGIKCHDIHCCTNLKCVREGSNRVCRKA SEQ ID NO: 341CAKKRNWCGKNEDCCCPMKCIYAWYNQQGSCQSTITGLFKKC SEQ ID NO: 342YCQKWMWTCDSARKCCEGLVCRLWCKKI SEQ ID NO: 343RGGCLPHNKFCNALSGPRCCSGLKCKELTIWNTKCLE SEQ ID NO: 344NVKCTGSKQCLPACKAAVGKAAGKCMNGKCKCYT SEQ ID NO: 345QRSCAKPGEMCMRIKCCDGQCGCNRGTGRCFCK SEQ ID NO: 346GCIPKHKRCTWSGPKCCNNISCHCNISGTLCKCRPG SEQ ID NO: 347NYCVAKRCRPGGRQCCSGKPCACVGKVCKCPRD SEQ ID NO: 348ERGCSGAYKRCSSSQRCCEGRPCVCSAINSNCKCRKT SEQ ID NO: 349RYCPRNPEACYNYCLRTGRPGGYCGGRSRITCFCFR SEQ ID NO: 350QRSCAKPGEMCMGIKCCDGQCGCNRGTGRCFCK SEQ ID NO: 351RRGCFKEGKWCPKSAPCCAPLKCKGPSIKQQKCVRE SEQ ID NO: 352TVKCGGCNRKCCAGGCRSGKCINGKCQCYGR SEQ ID NO: 353ERRCEPSGKPCRPLMRIPCCGSCVRGKCA SEQ ID NO: 354RGGCLPRNKFCNPSSGPRCCSGLTCKELNIWANKCL SEQ ID NO: 355CAKKRNWCGKNEDCCCPMKCIYAWYNQQGSCQTTITGLFKKC SEQ ID NO: 362VRIPVSCKHSGQCLKPCKDAGMRTGKCMNGKCDCTPK SEQ ID NO: 363VKCTTSKDCWPPCKKVTGRA SEQ ID NO: 364 GIVCRVCRIICGMQGRRVNICRAPIRCRCRRGSEQ ID NO: 365 SERDCIRHLQRCRENRDCCSRRCSRRGTNPERRCR SEQ ID NO: 366VRIPVSCRHSGQCLRPCRDAGMRFGRCMNGRCDCTPR SEQ ID NO: 367GVPINVRCRGSRDCLDPCRRAGMRFGRCINSRCHCTP SEQ ID NO: 368AVCVYRTCDRDCRRRGYRSGRCINNACRCYPYG SEQ ID NO: 369ISCTGSRQCYDPCRRRTGCPNARCMNRSCRCYGCG SEQ ID NO: 370QVQTNVRCQGGSCASVCRREIGVAAGRCINGRCVCYRN SEQ ID NO: 371EVIRCSGSRQCYGPCRQQTGCTNSRCMNRVCRCYGCG SEQ ID NO: 372ACRGVFDACTPGRNECCPNRVCSDRHRWCRWRL SEQ ID NO: 373QIYTSRECNGSSECYSHCEGITGRRSGRCINRRCYCYR SEQ ID NO: 374GCLEFWWRCNPNDDRCCRPRLRCSRLFRLCNFSFG SEQ ID NO: 375DCVRFWGRCSQTSDCCPHLACRSRWPRNICVWDGSVG SEQ ID NO: 376GCFGYRCDYYRGCCSGYVCSPTWRWCVRPGPGR SEQ ID NO: 377MNARFILLLVLTTMMLLPDTRGAEVIRCSGSRQCYGPCRQQTGCTNS RCMNRVCRCYGCGSEQ ID NO: 378 MNARLIYLLLVVTTMTLMFDTAQAVDIMCSGPRQCYGPCRRETGCPNARCMNRRCRCYGCV SEQ ID NO: 379MNARLIYLLLVVTTMMLTFDTTQAGDIRCSGTRQCWGPCRRQTTCTN SRCMNGRCRCYGCVGSEQ ID NO: 380 MNTRFIFLLLVVTNTMMLFDTRPVEGISCTGSRQCYDPCRRRTGCPNARCMNRSCRCYGCG SEQ ID NO: 381 GVPINVRCSGSRDCLEPCRRAGMRFGRCINRRCHCTPRSEQ ID NO: 382 GVPINVRCTGSPQCLRPCRDAGMRFGRCINGRCHCTPR SEQ ID NO: 383GVIINVRCRISRQCLEPCRRAGMRFGRCMNGRCHCTPR SEQ ID NO: 384GVPINVRCRGSPQCIQPCRDAGMRFGRCMNGRCHCTPQ SEQ ID NO: 385GVEINVRCTGSHQCIRPCRDAGMRFGRCINRRCHCTPR SEQ ID NO: 386GVEINVRCSGSPQCLRPCRDAGMRFGRCMNRRCHCTPR SEQ ID NO: 387GVPTDVRCRGSPQCIQPCRDAGMRFGRCMNGRCHCTPR SEQ ID NO: 388GVPINVSCTGSPQCIRPCRDAGMRFGRCMNRRCHCTPR SEQ ID NO: 389GVPINVPCTGSPQCIRPCRDAGMRFGRCMNRRCHCTPR SEQ ID NO: 390VGINVRCRHSGQCLRPCRDAGMRFGRCINGRCDCTPR SEQ ID NO: 391VGINVRCRHSGQCLRPCRDAGMRFGRCMNGRCDCTPR SEQ ID NO: 392VGIPVSCRHSGQCIRPCRDAGMRFGRCMNRRCDCTPR SEQ ID NO: 393RRGCFREGHSCPRTAPCCRPLVCRGPSPNTRRCTRP SEQ ID NO: 394SFCIPFRPCRSDENCCRRFRCRTTGIVRLCRW SEQ ID NO: 395LRGCLPRNRFCNALSGPRCCSGLRCRELSIWASRCL SEQ ID NO: 396GNYCLRGRCLPGGRRCCNGRPCECFARICSCRPR SEQ ID NO: 397TVRCGGCNRRCCPGGCRSGRCINGRCQCY SEQ ID NO: 398GCMREYCAGQCRGRVSQDYCLRHCRCIPR SEQ ID NO: 399ACLGFGERCNPSNDRCCRSSSLVCSQRHRWCRYG SEQ ID NO: 400RGGCLPHNRFCNALSGPRCCSGLRCRELSIRDSRCLG SEQ ID NO: 401RGGCLPRNRFCNPSSGPRCCSGLTCRELNIWASRCL SEQ ID NO: 402QRSCARPGDMCMGIRCCDGQCGCNRGTGRCFCR SEQ ID NO: 403ARGCADAYRSCNHPRTCCDGYNGYRRACICSGSNCRCRRS SEQ ID NO: 404RGGCLPHNRFCNALSGPRCCSGLRCRELSIWDSRCLG SEQ ID NO: 405RGGCLPHNRFCNALSGPRCCSGLRCRELSIYDSRCLG SEQ ID NO: 406RGGCLPHNRFCNALSGPRCCSRLRCRELSIWDSRCLG SEQ ID NO: 407RGGCLPHNRFCNALTGPRCCSRLRCRELSIWDSICLG SEQ ID NO: 408SCADAYKSCDSLRCCNNRTCMCSMIGTNCTCRRR SEQ ID NO: 409ERRCLPAGRTCVRGPMRVPCCGSCSQNRCT SEQ ID NO: 410LCSREGEFCYRLRRCCAGFYCRAFVLHCYRN SEQ ID NO: 411 ACGSCRRRCRGSGRCINGRCRCYSEQ ID NO: 412 ACGSCRRRCRGPGRCINGRCRCY SEQ ID NO: 413ACQGYMRRCGRDRPPCCRRLECSRTWRWCVWN SEQ ID NO: 414GRYCQRWMWTCDSRRACCEGLRCRLWCRRI SEQ ID NO: 415NARCRGSPECLPRCREAIGRAAGRCMNGRCRCYP SEQ ID NO: 416NVRCRGSRECLPACRAAVGRAAGRCMNGRCRCYP SEQ ID NO: 417NVRCRGSPECLPRCREAIGRSAGRCMNGRCRCYP SEQ ID NO: 418NARCRGSPECLPRCRQAIGRAAGRCMNGRCRCYP SEQ ID NO: 419RGYCAERGIRCHNIHCCSGLTCRCRGSSCVCRR SEQ ID NO: 420ERGCRLTFWRCRNRRECCGWNACALGICMPR SEQ ID NO: 421RRRCIARDYGRCRWGGTPCCRGRGCICSIMGTNCECRPR SEQ ID NO: 422GCRLTFWRCRNRRECCGWNACALGICMPR SEQ ID NO: 423ACRGLFVTCTPGRDECCPNHVCSSRHRWCRYR SEQ ID NO: 424IACAPRGLLCFRDRECCRGLTCRGRFVNTWPTFCLV SEQ ID NO: 425ACAGLYRRCGRGVNTCCENRPCRCDLAMGNCICRRR SEQ ID NO: 426FTCAISCDIRVNGRPCRGSGERRCSGGWSCRFNVCVRV SEQ ID NO: 427GFCAQRGIRCHDIHCCTNLRCVREGSNRVCRRA SEQ ID NO: 428CARRRNWCGRNEDCCCPMIRCIYAWYNQQGSCQSTITGLFRRC SEQ ID NO: 429YCQRWMWTCDSARRCCEGLVCRLWCRRI SEQ ID NO: 430RGGCLPHNRFCNALSGPRCCSGLRCRELTIWNTRCLE SEQ ID NO: 431NVRCTGSRQCLPACRAAVGRAAGRCMNGRCRCYT SEQ ID NO: 432QRSCARPGEMCMRIRCCDGQCGCNRGTGRCFCR SEQ ID NO: 433GCIPRHRRCTWSGPRCCNNISCHCNISGTLCRCRPG SEQ ID NO: 434NYCVARRCRPGGRQCCSGRPCACVGRVCRCPRD SEQ ID NO: 435ERGCSGAYRRCSSSQRCCEGRPCVCSAINSNCRCRRT SEQ ID NO: 436QRSCARPGEMCMGIRCCDGQCGCNRGTGRCFCR SEQ ID NO: 437RRGCFREGRWCPRSAPCCAPLRCRGPSIRQQRCVRE SEQ ID NO: 438TVRCGGCNRRCCAGGCRSGRCINGRCQCYGR SEQ ID NO: 439ERRCEPSGRPCRPLMRIPCCGSCVRGRCA SEQ ID NO: 440RGGCLPRNRFCNPSSGPRCCSGLTCRELNIWANRCL SEQ ID NO: 441CARRRNWCGRNEDCCCPMRCIYAWYNQQGSCQTTITGLFRRC SEQ ID NO: 448VRIPVSCRHSGQCLRPCRDAGMRTGRCMNGRCDCTPR SEQ ID NO: 451QKILSNRCNNSSECIPHCIRIFGTRAAKCINRKCYCYP SEQ ID NO: 452AVCNLKRCQLSCRSLGLLGKCIGDKCECVKHG SEQ ID NO: 453ISIGIRCSPSIDLCEGQCRIRRYFTGYCSGDTCHCSG SEQ ID NO: 454GDCLPHLRRCRENNDCCSRRCRRRGANPERRCR SEQ ID NO: 455SCEPGRTFRDRCNTCKCGADGRSAACTLRACPNQ SEQ ID NO: 456GDCLPHLKRCKADNDCCGKKCKRRGTNAEKRCR SEQ ID NO: 457GDCLPHLKRCKENNDCCSKKCKRRGTNPEKRCR SEQ ID NO: 458KDCLKKLKLCKENKDCCSKSCKRRGTNIEKRCR SEQ ID NO: 459GDCLPHLKRCKENNDCCSKKCKRRGANPEKRCR SEQ ID NO: 460VFINVKCRGSPECLPKCKEAIGKSAGKCMNGKCKCYP SEQ ID NO: 461VFINAKCRGSPECLPKCKEAIGKAAGKCMNGKCKCYP SEQ ID NO: 462VIINVKCKISRQCLEPCKKAGMRFGKCMNGKCHCTP SEQ ID NO: 463VPTDVKCRGSPQCIQPCKDAGMRFGKCMNGKCHCTP SEQ ID NO: 464VRIPVSCKHSGQCLKPCKDAGMRFGKCMNGKCDCTP SEQ ID NO: 465VRIPVSCRHSGQCLRPCRDAGMRFGRCMNGRCDCTP SEQ ID NO: 466TNVSCTTSKECWSVCQRLHNTSRGKCMNKKCRC SEQ ID NO: 467NVKCTGSKQCLPACKAAVGKAAGKCMNGKCKC SEQ ID NO: 468GVPINVRCRGSRDCLDPCRGAGERHGRCGNSRCHCTP SEQ ID NO: 469VRIPVSCRHSGQCLRPCRDAGERHGRCGGGRCDCTPR SEQ ID NO: 470QVQTNVRCQGGSCGSVCRREGGGAGGGCGNGRCGCYRN SEQ ID NO: 471GGDCLPHLRRCRADNDCCGRRCRRRGTNAERRCR SEQ ID NO: 472GGDCKYKFENWGACDGGTGTKVRQGTLKKARYNAQCQETIRVTKP C SEQ ID NO: 473GGDCKYKFENWGACDGGTGTKVRQGTLKKARYNAQCQETIRVTKP CTPKTKAKAKAKKGKGKDSEQ ID NO: 474 GGSCEPGRTFRDRCNTCRCGADGRSAACTLRACPNQ SEQ ID NO: 475GGQFTNVSCTTSRECWSVCQRLHNTSRGRCMNRRCRCYS SEQ ID NO: 476GGMCMPCFTTDHQMARRCDDCCGGRGRGRCYGPQCLCR SEQ ID NO: 477GGISIGIKCSPSIDLCEGQCRIRKYFTGYCSGDTCHCSG SEQ ID NO: 478GGEVIRCSGSKQCYGPCKQQTGCTNSKCMNKVCKCYGCG SEQ ID NO: 479GGSEKDCIKHLQRCRENKDCCSKKCSRRGTNPEKRCR SEQ ID NO: 480GGSCAKPRENCNRMNILCCRGECVCPTFGDCFCYGD SEQ ID NO: 481GGGVPINVKCRGSRDCLDPCKKAGMRFGKCINSKCHCTP SEQ ID NO: 482GGVRIPVSCKHSGQCLKPCKDAGMRFGKCMNGKCDCTPK SEQ ID NO: 483GGGIVCKVCKIICGMQGKKVNICKApIKCKCKKG SEQ ID NO: 484GGDCVRFWGKCSQTSDCCPHLACKSKWPRNICVWDGSVG SEQ ID NO: 485GGAVCVYRTCDKDCKRRGYRSGKCINNACKCYPYG SEQ ID NO: 486GGGCFGYKCDYYKGCCSGYVCSPTWKWCVRPGPGR SEQ ID NO: 487GGQVQTNVKCQGGSCASVCRREIGVAAGKCINGKCVCYRN SEQ ID NO: 488GGGDCLPHLKRCKENNDCCSKKCKRRGANPEKRCR SEQ ID NO: 489GGNFKVEGACSKPCRKYCIDKGARNGKCINGRCHCYY SEQ ID NO: 490GGQKILSNRCNNSSECIPHCIRIFGTRAAKCINRKCYCYP SEQ ID NO: 491GGDRDSCIDKSRCSKYGYYQECQDCCKKAGHNGGTCMFFKCKCA SEQ ID NO: 492GGAVCNLKRCQLSCRSLGLLGKCIGDKCECVKHG SEQ ID NO: 493GGQFCGTNGKPCVNGQCCGALRCVVTYHYADGVCLKMNP SEQ ID NO: 494GGRPTDIKCSASYQCFPVCKSRFGKTNGRCVNGLCDCF SEQ ID NO: 495GGNCAGYMRECKEKLCCSGYVCSSRWKWCVLPAPWRR SEQ ID NO: 496GGQFTDVKCTGSKQCWPVCKQMFGKPNGKCMNGKCRCYS SEQ ID NO: 497GGQIDTNVKCSGSSKCVKICIDRYNTRGAKCINGRCTCYP SEQ ID NO: 498GGAEIIRCSGTRECYAPCQKLTGCLNAKCMNKACKCYGCV SEQ ID NO: 499GGSDYCSNDFCFFSCRRDRCARGDCENGKCVCKNCHLN SEQ ID NO: 500GGCIGEGVPCDENDPRCCFGLVCLKPTLHGIWYKSYYCYKK SEQ ID NO: 501GGSCAKPGEMCMRIKCCDGQCGCNRGTGRCFCK SEQ ID NO: 502GGACLAEYQKCEGSTVPCCPGLSCSAGRFRKTKLCTK SEQ ID NO: 503GGVVIGQRCYRSPDCYSACKKLVGKATGKCTNGRCDC SEQ ID NO: 504GGACQFWSCNSSCISRGYRQGYCWGIQYKYCQCQ SEQ ID NO: 505GGRCPPCFTTNPNMEADCRKCCGGRGYCASYQCICPGG SEQ ID NO: 506GGVFINVKCRGSPECLPKCKEAIGKSAGKCMNGKCKCYP SEQ ID NO: 507GGQVSTNKKCSNTSQCYKTCEKVVGVAAGKCMNGKCICYP SEQ ID NO: 508GGECLEIFKACNPSNDQCCKSSKLVCSRKTRWCKYQIG SEQ ID NO: 509GGQDKCKKVYENYPVSKCQLANQCNYDCKLDKHARSGECFYDEKR NLQCICDYCEY SEQ ID NO: 510GGGHACYRNCWREGNDEETCKERC SEQ ID NO: 511GGMCNIPCFTTDTQMQERCDRCCGGGGRGRCWGPQCLCI SEQ ID NO: 512GGMCNIPCFTTEQRMAIICDDCCGGFGRGRCYGPQCLCR SEQ ID NO: 513GGICIPCFTTDHQIARRCDDCCGGRGRGRCYGPQCICR SEQ ID NO: 514GGRCQLQGFNCVVRSYGLPTIPCCRGLTCRSYFPGSTYGRCQRY SEQ ID NO: 515GGSFGLCRLRRGFCARGRCRFPSIPIGRCSRFVQCCRRVW SEQ ID NO: 516GGSCEPGTTFRDRCNTCRCGSDGRSAACTLRACPQ SEQ ID NO: 517GGSCTPGTTFRDRCNTCRCSSNGRSAACTLRACPPGSY SEQ ID NO: 518GGSCTPGTTFRNRCNTCRCGSNGRSASCTLMACPPGSY SEQ ID NO: 519GGSCTPGATFRNRCNTCRCGSNGRSASCTLMACPPGSY SEQ ID NO: 520GGSCQPGTTYQRGCNTCRCLEDGQTEACTLRLC SEQ ID NO: 521GGSCTPGATYREGCNICRCRSDGRSGACTRRICPVDSN SEQ ID NO: 522GGSCQPGTTFRRDCNTCVCNRDGTNAACTLRACL SEQ ID NO: 523GSYSRCQLQGFNCVVRSYGLPTIPCCRGLTCRSYFPGSTYGRCQRY SEQ ID NO: 524GGSCARPRENCNRMNILCCRGECVCPTFGDCFCYGD SEQ ID NO: 525GGGVPINVRCRGSRDCLDPCRRAGMRFGRCINSRCHCTP SEQ ID NO: 526GGSERDCIRHLQRCRENRDCCSRRCSRRGTNPERRCR SEQ ID NO: 527GGVRIPVSCRHSGQCLRPCRDAGMRFGRCMNGRCDCTPR SEQ ID NO: 528GGQVQTNVRCQGGSCASVCRREIGVAAGRCINGRCVCYRN SEQ ID NO: 529GGGDCLPHLRRCRENNDCCSRRCRRRGANPERRCR

In any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206,SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO:441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 529, or any fragmentthereof, any one or more K residues can be replaced by an R residue, anyone or more R residues can be replaced by a K residue or an A residue,any one or more A residues can be replaced by a K residue or an Rresidue, all K residues can be replaced by R residues or A residues, allbut one K residue can be replaced by R or A residues, all but two Kresidues can be replaced by R residues or A residues, or in anycombination thereof. In any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO:127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO:569, or any fragment thereof, any one or more M residues can be replacedby any one of I, L, or V residues, any one or more L residues can bereplaced by any one of V, I, or M residues, any one or more I residuescan be replaced by any one of M, L, or V residues, or any one or more Vresidues can be replaced by any one of I, L, or M residues. In anyembodiment, at least one of the amino acids alone or in combination canbe interchanged in the peptides or peptide fragments as follows: K/R,M/I L/V, G/A, S/T, Q/N, and D/E wherein each letter is each individuallyany amino acid or amino acid analogue. In some instances, the peptidecan contain only one lysine residue, or no lysine residue. In any of SEQID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213,SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, or SEQ ID NO: 451-SEQ ID NO: 569, or any fragment thereof, anyamino acid can be replaced with citrulline. In any of SEQ ID NO: 1-SEQID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, orSEQ ID NO: 451-SEQ ID NO: 569, or any fragment thereof, X canindependently be any number of any amino acid or no amino acid. In somecases, a peptide can include the first two N-terminal amino acids GS, aswith peptides of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO:206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 235, SEQ ID NO: 530-SEQID NO: 549, and SEQ ID NO: 570, a peptide can include the first twoN-terminal amino acids GG, as with peptides of SEQ ID NO: 471-SEQ ID NO:529, or such N-terminal amino acids (GS or GG) can be substituted by anyother one or two amino acids. In other cases, a peptide does not includethe first two N-terminal amino acids GS, as with peptides of SEQ ID NO:236-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 470, and SEQ ID NO: 550-SEQ ID NO: 569. In somecases, the N-terminus of the peptide is blocked, such as by an acetylgroup; in other instances the C-terminus of the peptide is block, suchas by an amide group.

In some instances, the peptide is any one of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO:451-SEQ ID NO: 569, or a functional fragment thereof. In otherembodiments, the peptide of the disclosure further comprises a peptidewith 100%, 99%, 97%, 95%, 90%, 85%, or 80% homology to any one of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, or SEQ ID NO: 451-SEQ ID NO: 569. In further embodiments, thepeptide fragment comprises a contiguous fragment of any one of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, or SEQ ID NO: 451-SEQ ID NO: 569 that is at least 17, at least 18,at least 19, at least 20, at least 21, at least 22, at least 23, atleast 24, at least 25, at least 26, at least 27, at least 28, at least29, at least 30, at least 31, at least 32, at least 33, at least 34, atleast 35, at least 36, at least 37, at least 38, at least 39, at least40, at least 41, at least 42, at least 43, at least 44, at least 45, atleast 46 residues long, wherein the peptide fragment is selected fromany portion of the peptide. In some embodiments, such peptide fragmentscontact the kidney and exhibit properties of those described herein forpeptide and peptide-active agent conjugates.

The peptides of the present disclosure can further comprise negativeamino acid residues. In some cases, the peptide has 2 or fewer negativeamino acid residues. In other cases, the peptide has 4 or fewer negativeamino acid residues, 3 or fewer negative amino acid residues, or 1 orfewer negative amino acid residues. The negative amino acid residues canbe selected from any negative charged amino acid residues. The negativeamino acid residues can selected from either E or D, or a combination ofboth E and D.

The peptides of the present disclosure can further comprise basic aminoacid residues. In some embodiments, basic residues are added to thepeptide sequence to increase the charge at physiological pH. The addedbasic residues can be any basic amino acid. The added basic residues canbe selected from K or R, or a combination of K or R.

In some embodiments, the peptide has a charge distribution comprising anacidic region and a basic region. An acidic region can be a nub. A nubis a portion of a peptide extending out of the peptide'sthree-dimensional structure. A basic region can be a patch. A patch is aportion of a peptide that does not designate any specific topologycharacteristic of the peptide's three-dimensional structure. In furtherembodiments, a cystine-dense peptide can be 6 or more basic residues and2 or fewer acidic residues.

The peptides of the present disclosure can further comprise positivelycharged amino acid residues. In some cases, the peptide has at least 2positively charged residues. In other cases, the peptide has at least 3positively charged residues, at least 4 positively charged residues, atleast 5 positively charged residues, at least 6 positively chargedresidues, at least 7 positively charged residues, at least 8 positivelycharged residues or at least 9 positively charged residues. Thepositively charged residues can be selected from any positively chargedamino acid residues. The positively charged residues can be selectedfrom either K or R, or a combination of K and R.

In addition, the peptides herein can comprise a 4-19 amino acid residuefragment of any of the above sequences containing at least 2 cysteineresidues, and at least 2 or 3 positively charged amino acid residues(for example, arginine, lysine or histidine, or any combination ofarginine, lysine or histidine). In other embodiments, the peptidesherein is a 20-70 amino acid residue fragment of any of the abovesequences containing at least 2 cysteine residues, no more than 2 basicresidues, and at least 2 or 3 positively charged amino acid residues(for example, arginine, lysine or histidine, or any combination ofarginine, lysine or histidine). In some embodiments, such peptidefragments contact the kidney and exhibit properties of those describedherein for peptide and peptide-active agent conjugates.

In some embodiments, the peptide contains one or more disulfide bondsand has a positive net charge at neutral pH. At physiological pH,peptides can have a net charge, for example, of −5, −4, −3, −2, −1, 0,+1, +2, +3, +4, or +5. When the net charge is zero, the peptide can beuncharged or zwitterionic. In some instances, the peptide can have apositive charge at physiological pH. In some instances, the peptide canhave a charge ≥+2 at physiological pH, ≥+3.5 at physiological pH, ≥+4.5at physiological pH. In some embodiments, the peptide contains one ormore disulfide bonds and has a positive net charge at neutral pH wherethe net charge can be +0.5 or less than +0.5, +1 or less than +1, +1.5or less than +1.5, +2 or less than +2, +2.5 or less than +2.5, +3 orless than +3, +3.5 or less than +3.5, +4 or less than +4, +4.5 or lessthan +4.5, +5 or less than +5, +5.5 or less than +5.5, +6 or less than+6, +6.5 or less than +6.5, +7 or less than +7, +7.5 or less than +7.5,+8 or less than +8, +8.5 or less than +8.5, +9 or less than +9.5, +10 orless than +10. In some embodiments, the peptide has a negative netcharge at physiological pH where the net charge can be −0.5 or less than−0.5, −1 or less than −1, −1.5 or less than −1.5, −2 or less than −2,−2.5 or less than −2.5, −3 or less than −3, −3.5 or less than −3.5, −4or less than −4, −4.5 or less than −4.5, −5 or less than −5, −5.5 orless than −5.5, −6 or less than −6, −6.5 or less than −6.5, −7 or lessthan −7, −7.5 or less than −7.5, −8 or less than −8, −8.5 or less than−8.5, −9 or less than −9.5, −10 or less than −10. In some cases, theengineering of one or more mutations within a peptide yields a peptidewith an altered isoelectric point, charge, surface charge, or rheologyat physiological pH. Such engineering of a mutation to a peptide derivedfrom a scorpion or spider can change the net charge of the complex, forexample, by decreasing the net charge by 1, 2, 3, 4, or 5, or byincreasing the net charge by 1, 2, 3, 4, or 5. In such cases, theengineered mutation may facilitate the ability of the peptide to contactthe kidney. Suitable amino acid modifications for improving the rheologyand potency of a peptide can include conservative or non-conservativemutations. A peptide can comprises at most 1 amino acid mutation, atmost 2 amino acid mutations, at most 3 amino acid mutations, at most 4amino acid mutations, at most 5 amino acid mutations, at most 6 aminoacid mutations, at most 7 amino acid mutations, at most 8 amino acidmutations, at most 9 amino acid mutations, at most 10 amino acidmutations, or another suitable number as compared to the sequence of thevenom or toxin that the peptide is derived from. In other cases, apeptide, or a functional fragment thereof, comprises at least 1 aminoacid mutation, at least 2 amino acid mutations, at least 3 amino acidmutations, at least 4 amino acid mutations, at least 5 amino acidmutations, at least 6 amino acid mutations, at least 7 amino acidmutations, at least 8 amino acid mutations, at least 9 amino acidmutations, at least 10 amino acid mutations, or another suitable numberas compared to the sequence of the venom or toxin that the peptide isderived from. In some embodiments, mutations can be engineered within apeptide to provide a peptide that has a desired charge or stability atphysiological pH.

Peptides can be mutated to add function or remove function. For example,peptides and peptide-conjugates of the present disclosure can be mutatedto retain, remove, or add the ability to bind to ion channels, or topromote agonizing or antagonizing ion channels, such as potassiumchannel binding that may occur with the peptide or peptide-conjugates(e.g., the potassium channel hERG). In some instances, it can beadvantageous to remove potassium channel binding from a peptide used fordelivery of an active agent. Mutations can include one or more N to Q, Nto A, N to S, N to T, N to L amino acid substitutions, or anycombination thereof, which can be made to decrease or eliminate peptidebinding to the ion channel. Alternatively, mutations can include one ormore S to G or S to R amino acid substitutions, or any combinationthereof, which can be made to retain function or to modulate functionduring peptide binding to the ion channel. In some embodiments thepeptides and peptide-drug conjugates of the present disclosure aremutated to minimize ion channel binding in order to minimize sideeffects or enhance the safety either in the target tissue orsystemically.

In some embodiments, charge can play a role in kidney homing. Positivelycharged residues can increase binding of peptides to proximal tubulecells, to megalin (which is negatively charged), or can otherwiseincrease retention in the kidney (Janzer et al. Bioconjug Chem. Oct. 4,2016, Geng et al. Bioconjug Chem. Jun. 20, 2012; 23(6):1200-10,Wischnjow et al. Bioconjug Chem. Apr. 20, 2016; 27(4):1050-7). Theinteraction of a peptide of this disclosure in solution and in vivo canbe influenced by the isoelectric point (pI) of the peptide and/or the pHof the solution or the local environment it is in. The charge of apeptide in solution can impact the solubility of the protein as well asparameters such as biodistribution, bioavailability, and overallpharmacokinetics. Additionally, positively charged molecules caninteract with negatively charged molecules. Positively charged moleculessuch as the peptides disclosed herein can interact and bind withmolecules such as megalin and cubilin, or another cell surface receptorexpressed by a cell of the proximal tubule, or a combination thereof.Positively charged residues can also interact with specific regions ofother proteins and molecules, such as negatively charged residues ofreceptors or electronegative regions of an ion channel pore on cellsurfaces. As such, the pI of a peptide can influence whether a peptideof this disclosure can efficiently home to the kidney. Identifying acorrelation between pI and kidney homing can be an important strategy inidentifying lead peptide candidates of the present disclosure. The pI ofa peptide can be calculated using a number of different methodsincluding the Expasy pI calculator and the Sillero method. The Expasy pIcan be determined by calculating pKa values of amino acids as describedin Bjellqvist et al., which were defined by examining polypeptidemigration between pH 4.5 to pH 7.3 in an immobilized pH gradient gelenvironment with 9.2M and 9.8M urea at 15° C. or 25° C. (Bjellqvist etal. Electrophoresis. 14(10):1023-31 (1993)). The Sillero method ofcalculating pI can involve the solution of a polynomial equation and theindividual pKas of each amino acid. This method does not use denaturingconditions (urea) (Sillero et al. 179(2): 319-35 (1989)) Using these pIcalculation methods and quantifying the kidney to blood ratio of peptidesignal after administration to a subject can be a strategy foridentifying a trend or correlation in charge and kidney homing. In someembodiments, a peptide with a pI above biological pH (˜pH 7.4) canexhibit efficient homing to kidney. In some embodiments, a peptide witha pI of at least 8, at least 9, at least 10, or at least 11 canefficiently home to kidney. In other embodiments, a peptide with a pI of11-12 can home most efficiently to kidney. In certain embodiments, apeptide can have a pI of about 9. In other embodiments, a peptide canhave a pI of 8-10. In some embodiments, more basic peptides can homemore efficiently to kidney. In other embodiments, a high pI alone maynot be sufficient to cause kidney homing of a peptide.

In some embodiments, the tertiary structure and electrostatics of apeptide of the disclosure can impact kidney homing. Structural analysisor analysis of charge distribution can be a strategy to predict residuesimportant in biological function, such as kidney homing. For example,several peptides of this disclosure that home to kidney can be groupedinto a structural class defined herein as “hitchins,” and can share theproperties of disulfide linkages between C1-C4, C2-C5, and C3-C6. Thefolding topologies of peptides knotted through three disulfide linkages(C1-C4, C2-C5, and C3-C6), can be broken down into structural familiesbased on the three-dimensional arrangement of the disulfides. Somecystine-dense peptides have the C3-C6 disulfide linkage passing throughthe macrocycle formed by the C1-C4 and C2-C5 disulfide linkages,hitchins have the C2-C5 disulfide linkage passing through the macrocycleformed by the C1-C4 and C3-C6 disulfide linkages, and yet otherstructural families have the C1-C4 disulfide linkage passing through themacrocycle formed by the C2-C5 and C3-C6 disulfide linkages. Variants of“hitchin” class peptides with preserved disulfide linkages at thesecysteine residues, primary sequence identity, and/or structural homologycan be a method of identifying or predicting other potential peptidecandidates that can home to kidney. Additionally, members and relatedmembers of the calcin family of peptides can also home to kidney,despite having a distinct tertiary structure from the “hitchin” class ofpeptides. Calcin peptides are structurally a subset of the cystine-densepeptides, with cystine-dense disulfide connectivity and topology, butare further classified on the basis of functioning to bind and activateryanodine receptors (RyRs). These receptors are calcium channels thatact to regulate the influx and efflux of calcium in muscle (Schwartz etal. Br J Pharmacol 157(3):392-403. (2009)). Variants of the calcinfamily of peptides with preserved key residues can be one way to predictpromising candidates that can home to kidney. In some embodiments,structural analysis of a peptide of this disclosure can be determined byevaluating peptides for resistance to degradation in buffers withvarious proteases or reducing agents. Structural analysis of thedistribution of charge density on the surface of a peptide can also be astrategy for predicting promising candidates that can home to kidney.Peptides with large patches of positive surface charge (when at pH 7.5)can home to kidney.

The NMR solution structures, x-ray crystallography, or crystalstructures of related structural homologs can be used to informmutational strategies that can improve the folding, stability, andmanufacturability, while maintaining the ability of a peptide to home tokidney. They can be used to predict the 3D pharmacophore of a group ofstructurally homologous scaffolds, as well as to predict possible graftregions of related proteins to create chimeras with improved properties.For example, this strategy can be used to identify critical amino acidpositions and loops that can be used to design drugs with improvedproperties or to correct deleterious mutations that complicate foldingand manufacturability for the peptides. These key amino acid positionsand loops can be retained while other residues in the peptide sequencescan be mutated to improve, change, remove, or otherwise modify function,homing, and activity of the peptide.

Additionally, the comparison of the primary sequences and the tertiarysequences of two or more peptides can be used to reveal sequence and 3Dfolding patterns that can be leveraged to improve the peptides and parseout the biological activity of these peptides. For example, comparingtwo different peptide scaffolds that home to kidney can lead to theidentification of conserved pharmacophores that can guide engineeringstrategies, such as designing variants with improved folding properties.Important pharmacophore, for example, can comprise aromatic residues orbasic residues, which can be important for binding.

Improved peptides can also be engineered based upon immunogenicityinformation, such as immunogenicity information predicted by TEPITOPEand TEPITOPEpan. TEPITOPE is a computational approach which usesposition specific scoring matrix to provide prediction rules for whethera peptide will bind to 51 different HLA-DR alleles, and TEPITOPEpan ismethod that uses TEPITOPE to extrapolate from HLA-DR molecules withknown binding specificities to HLA-DR molecules with unknown bindingspecificities based on pocket similarity. For example, TEPITOPE andTEPITOPEpan can be used to determine immunogenicity of peptides thathome to kidney. Immunogenicity information can also be predicted usingthe program NetMHCII version 2.3, which can determine the likelihoodthat a sequence might be presented as an immunogenic peptide via themajor histocompatibility complex (MEW) presentation system of antigenpresenting cells (APCs). (Nielson, M et al. BMC Bioinformatics, 8: 238(2007); Nielsen, M. et al. BMC Bioinformatics, 10: 296 (2009)). Thisprogram can create an immunogenicity score by predicting the binding ofa peptide to MEW alleles. Strong binding alleles and weak bindingalleles in each major MEW allele group (DR, DQ, and DP) can be talliedseparately. The number of peptides of a specific length within thesequence (e.g., a ‘core’ peptide that can be nine residues long) thatare immunogenic can also be tallied. Comparison of peptides or ‘core’peptides with high immunogenecity to peptides or ‘core’ peptides withlow immunogenicity can guide engineering strategies for designingvariants with decreased immunogenicity. Stronger binding peptides can bemore likely to generate an immune response in patient carrying thatgiven MEW alleles. Mutating stronger binding amino acids or peptides outof a peptide sequence can reduce the immunogenicity of the entirepeptide. Another aspect of immunogenicity, in addition to whether apeptide binds to a patient's MEW allele, can be whether the patient'simmune cells, such as a professional antigen presenting cells such as amacrophage, a B cell, or a dendritic cell, can process the peptide. Adendritic cell can take up a protein or peptide, and then can process apeptide, such as by cleaving to form a nine residue long peptide, whichthen can bind to the MEW and can be presented on the surface of thedendritic cell to the immune system's various T cells, including helperT cells and cytotoxic T cells, and thus can stimulate an immuneresponse. The processing can involve peptide bond cleavage by enzymesand disulfide bond reduction, and thus a peptide or protein that isresistant to enzymatic cleavage and/or reduction can be resistant toprocessing and subsequent MEW presentation to the immune system.Therefore, having a peptide or protein that is resistant to enzymaticcleavage and/or reduction can reduce its immunogenic potential.

In some embodiments, a peptide of this disclosure can bind to interactwith, modulate, antagonize, or agonize any of the below renal ionchannels in TABLE 2 reproduced from Table 1 of Kuo et al. (Chem Rev.2012 Dec. 12; 112(12): 6353-6372), which is incorporated herein byreference.

TABLE 2 Renal Ion Channels from Kuo et al. Protein (gene) nameDistribution in kidney Ion affected Disease associated TRPC6 (Trpc6)Glomerulus Ca²⁺ Focal Segmental Glomerulosclerosis TRPM6 (Trpm6) Distalconvoluted tubule Mg²⁺ Hypomagnesemia ClC-5 (CLCN5) Convoluted proximaltubule Cl⁻/H⁺ Dent's disease ClC-Kb (CLCNKB) Thick ascending loop of Cl⁻Bartter syndrome Henle ROMK (KCNJ1) Thick ascending loop of K⁺ Barttersyndrome Henle; Distal nephron Kir4.1 (KCNJ10) Collecting duct K⁺ EASTsyndrome ENaC (Scnn1a) Collecting duct Na⁺ Pseudohypoaldosteronism ENaC(Scnn1a) Collecting duct Na⁺ Liddle's syndrome Polycystin 2 (PKD2)Convoluted tubule Ca²⁺ Polycystic kidney disease

In some embodiments, a peptide of this disclosure can bind to, inter actwith, modulate antagonize, or agonize any of the below renal ionchannels in TABLE 3 reproduced from Table 1 of Zhou et al. (Am J PhysiolRenal Physiol. Jun. 1, 2016; 310(11): F1157-F1167), which isincorporated herein by reference.

TABLE 3 Renal Ion Channels from Zhou et al. Protein Human GeneExpression in Agonists and Function in the Relevant Kidney Name Name theKidney Activators Kidney Diseases TRP family TRPC TRPC1 TRPC1, TRP1 MCPLC Regulates DN mesangial cell contractility TRPC3 TRPC3, TRP3 P, DCT,CD DAG, PLC Regulates SOCE Williams-Beuren in podocytes, syndrome Ca²⁺reabsorption hypercalcemia, in DCT and renal fibrosis CD TRPC5 TRPC5,TRP5 P, JGC Intracellular Dysregulates Podocyte injury, Ca²⁺, podocyteactin glomerular disease lysophospholipids, cytoskeleton, oxidativedegrades stress, synaptopodin, rosiglitazone, and activates riluzole,PLC Rac1 TRPC6 TRPC6, TRP6, P, CD PLC, DAG, Regulates FSGS, DN FSGS2hyperforin, podocyte slit lysophosphatidyl- diaphragm choline, 20-HETETRPV TRPV4 TRPV4, VR- ATL, TAL, Mechanical Regulates renal OAC, OTRPCDCT, CNT stress, warm osmolality and 4 (<33° C.), 4α- water PDD,reabsorption GSK1016790 A TRPV5 TRPV5, CAT2, DCT, CNT ConstitutivelyCa²⁺ reabsorption ECaC1 active, PKA- dependent phosphorylation, sheerstress, PIP₂ TRPP PKD1/PKD2 PKD1/PKD2 Epithelial Mechanical Activates GADPKD complex cells of TAL, stress, protein signaling DCT intracellularcascades, Ca²⁺ (?) mechanosensor VGCC T-type VGCC Ca_(v)3.1 CACNA1GAfferent and Low voltage Regulates blood DN, fibrosis, efferent flowglomerular arterioles, hypertension MC, DCT, CD Ca_(v)3.2 CACNA1HAfferent and Low voltage Regulates efferent glomerular arterioles,filtration rate MC, L-type VGCC Ca_(v)1.2 CACNA1C Afferent and Highvoltage, Vasoconstriction, Glomerular efferent 1,4- modifies thehypertension, arterioles, dihydropyridines, formation of PKD (?) MC, DCTFPL- kidney cysts 64176 P-/Q-type VGCC Ca_(v)2.1 CACNA1A Afferent Highvoltage Depolarization- arterioles, MC mediated contraction in renalafferent arterioles TRP, transient receptor potential; VGCC,voltage-gated calcium channels; P, podocyte; MC, mesangial cell; PCT,proximal convoluted tubule; ATL, ascending thin limb; TAL, thickascending limb; DCT, distal convoluted tubule; CNT, connecting tubule;CD, collecting duct; SOCE, store-operated Ca2+ entry; PIP₂,phosphatidylinositol 4,5-bisphosphate; JGC, juxtaglomerular cell; ADPKD,autosomal dominant polycystic kidney disease; DN, diabetic nephropathy;NDI, nephrogenic diabetes insipidus; FSGS, focal segmentalglomerulosclerosis.

In some embodiments, a peptide of this disclosure can bind to, inter actwith, modulate antagonize, or agonize any of the renal ion channels inLoudon et al. (Ann Clin Biochem. 2014 July; 51(Pt 4):441-58), which isincorporated herein by reference. Such renal ion channels include NKCC2,ROMK, ClC-Kb, ClC-Ka, NCCT, TRPM6, TRPM7, Kv1.1, Kir4.1, ROMK1, Maxi-K,ENaC, PC1, PC2, and CLC-5, or any combination thereof.

Furthermore, multiple sequence alignment can also be used to informmutational strategies using previously identified sequences, and thusproviding a guide to making changes that would eliminate labile residuesand immunogenic regions of a peptide sequence. Peptides can be evaluatedfor residues of potential biochemical instability and regions ofpotential immunogenicity. Then, a residue that can allow for greaterpeptide stability at a certain location in a peptide can be identifiedfrom a multiple sequence alignment. For example, a specific residue canbe identified from a multiple sequence alignment as providing greaterstability for a peptide at position previously identified as a possiblerisk for a significant rate of deamidation, cleavage, degradation,oxidation, hydrolysis, isomerization, disulfide exchange, racemization,beta elimination, or aggregation. This information can then be used tocreate peptides with greater stability or reduced immunogenicity.

In addition to utilizing co-crystal x-ray structures, NMR solutionstructures, and mutagenesis studies, a multiple alignment of peptidesequences can be used to identify specific amino acids or regions ofhigh conservation that indicate an important interaction with a targetor receptor (e.g., binding to a potassium channel protein) or areimportant for folding and structure or other properties. Once theconserved amino acid or region is identified, then amino acidsreplacements can be determined that maintain the important properties ofthe peptide, such as maintenance of the structure, reduction inimmunogenicity, reduction in binding to an ion channel protein,increased stability, or any combination of thereof.

The multiple sequence alignment can also identify possible locations toadd a tyrosine or tryptophan residue for spectrophotometric reporting.Incorporation of aromatic amino acids such as Tyrosine or Tryptophaninto a peptide such as SEQ ID NO: 132, which otherwise contains onlyamino acids of low UV absorbance at 280 nm, can be analyticallyadvantageous. Tyrosine and Tryptophan amino acids contain aromatic ringstructures. These residues have distinct absorption and emissionwavelengths and good quantum yields, as shown in TABLE 4 not present inother amino acids. Both Tyrosine and Tryptophan can provide a good‘handle’ for analytical detection of a peptide in solution since UVabsorbance in the 250-300 nm range and peptide fluorescence is specificfor these aromatic molecules. While detection of a peptide such as SEQID NO: 132 relies on the absorbance of the peptide bond at 220 nm, wheremany other materials including minor impurities in solvents also oftencontribute to signal, the absorbance and fluorescence properties ofTryptophan and Tyrosine containing peptides can provide for asignificantly more selective and sensitive detection. Thus incorporatingan aromatic amino acid can create peptides better suited forconcentration and purity measurements, which can be useful duringanalytics, process development, manufacturing, and other drugdevelopment and drug manufacturing activities. Incorporation can beachieved either through substitutions of one or more amino acids in thepeptide to Tyr and/or Trp, insertion of Tyr and/or Trp into the peptide,or via addition of Tyr and/or Trp to the N-terminus or C-terminus of thepeptide.

TABLE 4 Absorbance and Fluorescence Characteristics of Tryptophan andTyrosine. Absorbance Fluorescence Amino Wavelength AbsorbtivityWavelength Quantum Acid (nm) (M*cm)⁻¹ (nm) Yield Tryptophan 280 5,600348 0.20 Tyrosine 274 1,400 303 0.14

A peptide of this disclosure can bind to chloride, potassium, or sodiumchannels. The peptide can also bind to calcium or magnesium channels.The peptide can block potassium channels and/or sodium channels. In someembodiments, the peptide can block any one or more of such channels. Insome embodiments, the peptide cannot interact with any of such channelsor can be mutated to reduce or remove binding to any such channels. Thepeptide can block calcium or magnesium channels. In some embodiments,the peptide can activate any one or more of such channels. In stillother embodiments, the peptide can be a potassium channel agonist, apotassium channel antagonist, a portion of a potassium channel, a sodiumchannel agonist, a sodium channel antagonist, a chloride channelagonist, a chloride channel antagonist, a calcium channel agonist, acalcium channel antagonist, a hadrucalcin, a theraphotoxin, ahuwentoxin, a kaliotoxin, a cobatoxin, or a lectin. In some embodiments,the lectin can be SHL-Ib2. In some embodiments, the peptide can interactwith, binds, inhibits, inactivates, or alters expression of ion channelsor chloride channels. In some embodiments, the peptide can interact withan Nav1.7 ion channel. In some embodiments, the peptide can interactwith a Kv 1.3 ion channel. In still other embodiments, the peptideinteracts with proteases, matrix metalloproteinase, inhibits cancer cellmigration or metastases, has antimicrobial activity, or has antitumoractivity. In addition to acting on matrix metalloproteinases, thepeptide can interact with other possible proteases (e.g., elastases). Insome embodiments, a peptide of this disclosure can bind to multidrugresistance transporters. Peptide and peptide drug conjugate binding toand blocking multidrug resistance transporters can be used to treatbacterial infections or cancers of the kidney.

The present disclosure can also encompass multimers of the variouspeptides described herein. Examples of multimers include dimers,trimers, tetramers, pentamers, hexamers, heptamers, and so on. Amultimer can be a homomer formed from a plurality of identical subunitsor a heteromer formed from a plurality of different subunits. In someembodiments, a peptide of the present disclosure is arranged in amultimeric structure with at least one other peptide, or two, three,four, five, six, seven, eight, nine, ten, or more other peptides. Incertain embodiments, the peptides of a multimeric structure each havethe same sequence. In alternative embodiments, some or all of thepeptides of a multimeric structure have different sequences.

The present disclosure further includes peptide scaffolds that, e.g.,can be used as a starting point for generating additional peptides. Insome embodiments, these scaffolds can be derived from a variety ofcystine-dense peptides. Some suitable peptides for scaffolds caninclude, but are not limited to, chlorotoxin, brazzein, circulin,stecrisp, hanatoxin, midkine, hefutoxin, potato carboxypeptidaseinhibitor, bubble protein, attractin, α-GI, α-GID, μ-PIIIA, ω-MVIIA,ω-CVID, χ-MrIA, ρ-TIA, conantokin G, contulakin G, GsMTx4, margatoxin,shK, toxin K, chymotrypsin inhibitor (CTI), and EGF epiregulin core.

In some embodiments, the peptide sequences of the disclosure are flankedby additional amino acids. One or more additional amino acids can, forexample, confer a desired in vivo charge, isoelectric point, chemicalconjugation site, stability, or physiologic property to a peptide.

Identifying sequence homology can be important for determining keyresidues that preserve kidney targeting function. For example, in someembodiments retention of conserved hydrophilic residues, such as N, Q,S, T, D, E, K, R, and H, can be important in preserving peptide kidneytargeting function by keeping the peptide from sticking to albumin andsuch function also engineered into any variants that are made. In otherembodiments, identification of basic amino acids such as Lys and/or Argcan important to binding and retention of a peptide in the kidney andsuch function also engineered into any variants that are made. Two ormore peptides can share a degree of homology and share similarproperties in vivo. For instance, a peptide can share a degree ofhomology with a peptide of the present disclosure. In some cases, apeptide of the disclosure can have up to about 20% pairwise homology, upto about 25% pairwise homology, up to about 30% pairwise homology, up toabout 35% pairwise homology, up to about 40% pairwise homology, up toabout 45% pairwise homology, up to about 50% pairwise homology, up toabout 55% pairwise homology, up to about 60% pairwise homology, up toabout 65% pairwise homology, up to about 70% pairwise homology, up toabout 75% pairwise homology, up to about 80% pairwise homology, up toabout 85% pairwise homology, up to about 90% pairwise homology, up toabout 95% pairwise homology, up to about 96% pairwise homology, up toabout 97% pairwise homology, up to about 98% pairwise homology, up toabout 99% pairwise homology, up to about 99.5% pairwise homology, or upto about 99.9% pairwise homology with a second peptide. In some cases, apeptide of the disclosure can have at least about 20% pairwise homology,at least about 25% pairwise homology, at least about 30% pairwisehomology, at least about 35% pairwise homology, at least about 40%pairwise homology, at least about 45% pairwise homology, at least about50% pairwise homology, at least about 55% pairwise homology, at leastabout 60% pairwise homology, at least about 65% pairwise homology, atleast about 70% pairwise homology, at least about 75% pairwise homology,at least about 80% pairwise homology, at least about 85% pairwisehomology, at least about 90% pairwise homology, at least about 95%pairwise homology, at least about 96% pairwise homology, at least about97% pairwise homology, at least about 98% pairwise homology, at leastabout 99% pairwise homology, at least about 99.5% pairwise homology, atleast about 99.9% pairwise homology with a second peptide. Variousmethods and software programs can be used to determine the homologybetween two or more peptides, such as NCBI BLAST, Clustal W, MAFFT,Clustal Omega, AlignMe, Praline, or another suitable method oralgorithm.

In still other instances, the variant nucleic acid molecules encoding apeptide of any one of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ IDNO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 529 canbe identified by either a determination of the sequence identity orhomology of the encoded peptide amino acid sequence with the amino acidsequence of any one of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 529, orby a nucleic acid hybridization assay. Such peptide variants can includenucleic acid molecules (1) that remain hybridized with a nucleic acidmolecule having the nucleotide sequence encoded by any one of SEQ ID NO:1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ IDNO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448,or SEQ ID NO: 451-SEQ ID NO: 529 (or any complement of the previoussequences) under stringent washing conditions, in which the washstringency is equivalent to 0.5×−2×SSC with 0.1% SDS at 55-65° C., and(2) that encode a peptide having at least 70%, at least 80%, at least90%, at least 95% or greater than 95% sequence identity or homology tothe amino acid sequence of any one SEQ ID NO: 1-SEQ ID NO: 120, SEQ IDNO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355,SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ IDNO: 529. Alternatively, peptide variants of any one of SEQ ID NO: 1-SEQID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, orSEQ ID NO: 451-SEQ ID NO: 529 can be characterized as nucleic acidmolecules (1) that remain hybridized with a nucleic acid molecule havingthe nucleotide sequence encoding any one of SEQ ID NO: 1-SEQ ID NO: 120,SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO:355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO:451-SEQ ID NO: 529 (or any complement of the previous sequences) underhighly stringent washing conditions, in which the wash stringency isequivalent to 0.1×−0.2×SSC with 0.1% SDS at 50-65° C., and (2) thatencode a peptide having at least 70%, at least 80%, at least 90%, atleast 95% or greater than 95% sequence identity or homology to the aminoacid sequence of any one of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO:127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO:529.

Percent sequence identity or homology can be determined by conventionalmethods. See, for example, Altschul et al., Bull. Math. Bio. 48:603(1986), and Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89:10915(1992). Briefly, two amino acid sequences are aligned to optimize thealignment scores using a gap opening penalty of 10, a gap extensionpenalty of 1, and the “BLOSUM62” scoring matrix of Henikoff and Henikoff(Id.). The sequence identity or homology is then calculated as: ([Totalnumber of identical matches]/[length of the longer sequence plus thenumber of gaps introduced into the longer sequence in order to align thetwo sequences])(100).

Additionally, there are many established algorithms available to aligntwo amino acid sequences. For example, the “FASTA” similarity searchalgorithm of Pearson and Lipman is a suitable protein alignment methodfor examining the level of sequence identity or homology shared by anamino acid sequence of a peptide disclosed herein and the amino acidsequence of a peptide variant. The FASTA algorithm is described byPearson and Lipman, Proc. Nat'l Acad. Sci. USA 85:2444 (1988), and byPearson, Meth. Enzymol. 183:63 (1990). Briefly, FASTA firstcharacterizes sequence similarity by identifying regions shared by thequery sequence (e.g., SEQ ID NO: 530) and a test sequence that haseither the highest density of identities (if the ktup variable is 1) orpairs of identities (if ktup=2), without considering conservative aminoacid substitutions, insertions, or deletions. The ten regions with thehighest density of identities are then rescored by comparing thesimilarity of all paired amino acids using an amino acid substitutionmatrix, and the ends of the regions are “trimmed” to include only thoseresidues that contribute to the highest score. If there are severalregions with scores greater than the “cutoff” value (calculated by apredetermined formula based upon the length of the sequence and the ktupvalue), then the trimmed initial regions are examined to determinewhether the regions can be joined to form an approximate alignment withgaps. Finally, the highest scoring regions of the two amino acidsequences are aligned using a modification of theNeedleman-Wunsch-Sellers algorithm (Needleman and Wunsch, J. Mol. Biol.48:444 (1970); Sellers, Siam J. Appl. Math. 26:787 (1974)), which allowsfor amino acid insertions and deletions. Illustrative parameters forFASTA analysis are: ktup=1, gap opening penalty=10, gap extensionpenalty=1, and substitution matrix=BLOSUM62. These parameters can beintroduced into a FASTA program by modifying the scoring matrix file(“SMATRIX”), as explained in Appendix 2 of Pearson, Meth. Enzymol.183:63 (1990).

FASTA can also be used to determine the sequence identity or homology ofnucleic acid molecules using a ratio as disclosed above. For nucleotidesequence comparisons, the ktup value can range between one to six,preferably from three to six, most preferably three, with otherparameters set as described above.

Some examples of common amino acids that are a “conservative amino acidsubstitution” are illustrated by a substitution among amino acids withineach of the following groups: (1) glycine, alanine, valine, leucine, andisoleucine, (2) phenylalanine, tyrosine, and tryptophan, (3) serine andthreonine, (4) aspartate and glutamate, (5) glutamine and asparagine,and (6) lysine, arginine and histidine. The BLOSUM62 table is an aminoacid substitution matrix derived from about 2,000 local multiplealignments of protein sequence segments, representing highly conservedregions of more than 500 groups of related proteins (Henikoff andHenikoff, Proc. Nat'l Acad. Sci. USA 89:10915 (1992)). Accordingly, theBLOSUM62 substitution frequencies can be used to define conservativeamino acid substitutions that may be introduced into the amino acidsequences of the present invention. Although it is possible to designamino acid substitutions based solely upon chemical properties (asdiscussed above), the language “conservative amino acid substitution”preferably refers to a substitution represented by a BLOSUM62 value ofgreater than −1. For example, an amino acid substitution is conservativeif the substitution is characterized by a BLOSUM62 value of 0, 1, 2, or3. According to this system, preferred conservative amino acidsubstitutions are characterized by a BLOSUM62 value of at least 1 (e.g.,1, 2 or 3), while more preferred conservative amino acid substitutionsare characterized by a BLOSUM62 value of at least 2 (e.g., 2 or 3).

Determination of amino acid residues that are within regions or domainsthat are critical to maintaining structural integrity can be determined.Within these regions one can determine specific residues that can bemore or less tolerant of change and maintain the overall tertiarystructure of the molecule. Methods for analyzing sequence structureinclude, but are not limited to, alignment of multiple sequences withhigh amino acid or nucleotide identity or homology and computer analysisusing available software (e.g., the Insight II™ viewer and homologymodeling tools; MSI, San Diego, Calif.), secondary structurepropensities, binary patterns, complementary packing and buried polarinteractions (Barton, G. J., Current Opin. Struct. Biol. 5:372-6 (1995)and Cordes, M. H. et al., Current Opin. Struct. Biol. 6:3-10 (1996)). Ingeneral, when designing modifications to molecules or identifyingspecific fragments determination of structure can typically beaccompanied by evaluating activity of modified molecules.

Pairwise sequence alignment is used to identify regions of similaritythat may indicate functional, structural and/or evolutionaryrelationships between two biological sequences (protein or nucleicacid). By contrast, multiple sequence alignment (MSA) is the alignmentof three or more biological sequences. From the output of MSAapplications, homology can be inferred and the evolutionary relationshipbetween the sequences assessed. One of skill in the art would recognizeas used herein, “sequence homology” and “sequence identity” and “percent(%) sequence identity” and “percent (%) sequence homology” have beenused interchangeably to mean the sequence relatedness or variation, asappropriate, to a reference polynucleotide or amino acid sequence.

In some embodiments, the first two N-terminal amino acids of SEQ ID NO:1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ IDNO: 216-SEQ ID NO: 235, SEQ ID NO: 471-SEQ ID NO: 529, SEQ ID NO:530-SEQ ID NO: 549, or SEQ ID NO: 570 (GS or GG) serve as a spacer orlinker in order to facilitate conjugation or fusion to another molecule,as well as to facilitate cleavage of the peptide from such conjugated orfused molecules. In some embodiments, the peptide may not include thefirst two N-terminal amino acids shown in SEQ ID NO: 236-SEQ ID NO: 355,SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO:470 or SEQ ID NO: 550-SEQ ID NO: 569, or such N-terminal amino acids canbe substituted by any other one or two amino acids, as shown in SEQ IDNO: 471-SEQ ID NO: 529. For example, in certain embodiments, the firsttwo N-terminal amino acids (GS) of SEQ ID NO: 1 are substituted with GGas in SEQ ID NO: 471. As another example, in certain embodiments, thefirst two N-terminal amino acids (GG) of SEQ ID NO: 40 are substitutedwith GS as in SEQ ID NO: 523.

In some embodiments, the peptide sequence is flanked by additional aminoacids. One or more additional amino acids can, for example, confer adesired charge under physiological conditions, isoelectric point,chemical conjugation site, stability, or physiologic property to apeptide. For instance, the amine in a lysine residue or the N-terminuscan serve as a chemical conjugation site. Other lysine residues can bemutated out, such as by substitution with arginine, to provide a singlesite for amine conjugation.

The present disclosure encompasses various modifications to the peptidesprovided herein. In some embodiments, a peptide of the presentdisclosure contains or is modified to contain only one lysine residue,or no lysine residues. In some embodiments, some or all of the lysineresidues in the peptide are replaced with arginine residues. In someembodiments, some or all of the methionine residues in the peptide arereplaced by leucine or isoleucine. In some embodiments, some or all ofthe tryptophan residues in the peptide are replaced by phenylalanine ortyrosine. In some embodiments, some or all of the asparagine residues inthe peptide are replaced by glutamine. In some embodiments, some or allof the cysteine residues in the peptide are replaced by serine toproduce a linearized form of the peptide. In some embodiments, theN-terminus of the peptide is blocked, such as by an acetyl group. Insome embodiments, the N-terminus of the peptide is blocked withpyroglutamic acid. Alternatively or in combination, in some instances,the C-terminus of the peptide is blocked, such as by an amide group. Insome embodiments, the peptide is modified by methylation on free amines.For example, full methylation can be accomplished through the use ofreductive methylation with formaldehyde and sodium cyanoborohydride.

At physiological pH, peptides can have a net charge, for example, of −5,−4, −3, −2, −1, 0, +1, +2, +3, +4, +5, +6, +7, +8, +9, or +10. When thenet charge is zero, the peptide can be uncharged or zwitterionic. Insome embodiments, the engineering of one or more mutations within apeptide yields a peptide with an altered isoelectric point, charge,surface charge, or rheology at physiological pH. Such engineering of amutation to a peptide of the present disclosure (e.g., a peptide derivedfrom a scorpion or spider) can change the net charge of the complex, forexample, by decreasing the net charge by 1, 2, 3, 4, or 5, or byincreasing the net charge by 1, 2, 3, 4, or 5.

In certain embodiments, the engineered mutation can facilitate theability of the peptide to bind to renal tissue. Suitable amino acidmodifications for improving the rheology and potency of a peptide caninclude conservative or non-conservative mutations. A peptide cancomprise at most 1 amino acid mutation, at most 2 amino acid mutations,at most 3 amino acid mutations, at most 4 amino acid mutations, at most5 amino acid mutations, at most 6 amino acid mutations, at most 7 aminoacid mutations, at most 8 amino acid mutations, at most 9 amino acidmutations, at most 10 amino acid mutations, or another suitable numberas compared to the sequence of the peptide scaffold (e.g., venom ortoxin component) that the peptide is derived from. In other cases, apeptide, or a functional fragment thereof, comprises at least 1 aminoacid mutation, at least 2 amino acid mutations, at least 3 amino acidmutations, at least 4 amino acid mutations, at least 5 amino acidmutations, at least 6 amino acid mutations, at least 7 amino acidmutations, at least 8 amino acid mutations, at least 9 amino acidmutations, at least 10 amino acid mutations, or another suitable numberas compared to the sequence of the peptide scaffold (e.g., venom ortoxin component) that the peptide is derived from. In some embodiments,mutations can be engineered within a peptide to provide a peptide thathas a desired charge or stability at physiological pH.

In some embodiments, more than one peptide sequence is present on aparticular peptide. For example, a peptide of the present disclosure caninclude sequences from at least 1, at least 2, at least 3, at least 4,at least 5, at least 6, at least 7, at least 8, at least 9, or at least10 different peptides, or fragments thereof.

Peptide Properties for Renal Localization, Binding, and Internalization

The present disclosure provides peptides that can distribute to, home,target, be directed to, accumulate in, migrate to, be retained in,and/or bind to one or more specific regions, tissue, structures,regions, compartments, or cells of the kidney, collectively referred toherein as “renal tissue.” Examples of regions, tissue, structures, orcells of the kidney applicable to the embodiments presented hereininclude but are not limited to: the cortex region, the glomerulus, theglomerular filtrate (Bowman's space) tubular lumina, the proximaltubule, the S1, S2, and S3 segments, the medulla region, the descendingtubule, the ascending tubule, the distal tubule, the loop of Henle, theBowman's capsule, the renal interstitium, the renal microvasculature,vasa rectae, or any cells or cell types thereof.

In some embodiments, the peptides of the present disclosure interactwith renal tissue of the subject, e.g., by binding to the renal tissue.The binding between the peptide and the renal tissue can be a specificbinding interaction (e.g., a receptor-ligand interaction) ornon-specific binding interaction (e.g., electrostatic interaction). Forexample, in certain embodiments, upon administration to a subject, apeptide of the present disclosure binds to a proximal tubule of thesubject, e.g., a cell of the proximal tubule. As another example, incertain embodiments, upon administration to a subject, a peptide of thepresent disclosure binds to a glomerulus of the subject, e.g., a cell ofthe glomerulus. As another example, in certain embodiments, a peptide ofthe present disclosure binds to podocytes. In various embodiments, thepeptides bind to receptors expressed by a renal cell. For instance, apeptide can bind to a cell surface receptor expressed by a cell of theproximal tubule, a megalin receptor, a cubulin receptor, or acombination thereof.

In some embodiments, the peptides are internalized by a cell of therenal tissue of the subject. The present disclosure encompasses varioustypes of internalization mechanisms, including but not limited topinocytosis, phagocytosis, endocytosis, receptor-mediated endocytosis,scavenging mechanisms, membrane penetration or translocation mechanisms,or combinations thereof. For example, a peptide can be internalizedfollowing binding to the cell or a receptor thereof, e.g., viareceptor-mediated endocytosis.

Certain embodiments of the peptides described herein exhibit propertiesthat enhance localization, binding, accumulation in, and/orinternalization by renal tissues, regions, compartments, or cells.Examples of peptide properties that can be relevant to renal binding andinternalization include but are not limited to isoelectric point, netcharge, charge distribution, molecular weight, hydrodynamic radius, pHstability, hydrophilicity, and protein-protein binding.

For example, in various embodiments, the peptides of the presentdisclosure exhibit an isoelectric point (pI) favorable for renallocalization, binding, and/or internalization. In certain embodiments,the pI of a peptide is less than or equal to about 2.0, less than orequal to about 2.5, less than or equal to about 3.0, less than or equalto about 3.5, 4.0, less than or equal to about 4.5, less than or equalto about 5.5, less than or equal to about 6.0, less than or equal toabout 6.5, less than or equal to about 7.0, less than or equal to about7.5, less than or equal to about 8.0, less than or equal to about 8.5,less than or equal to about 9.0, less than or equal to about 9.5, lessthan or equal to about 10.0, less than or equal to about 10.5, less thanor equal to about 11.0, less than or equal to about 11.5, less than orequal to about 12.0, less than or equal to about 12.5, less than orequal to about 13.0, less than or equal to about 13.5, less than orequal to about 14.0, less than or equal to about 14.5, or less than orequal to about 15.0. In certain embodiments, the pI of a peptide isgreater than or equal to about 2.0, greater than or equal to about 2.5,greater than or equal to about 3.0, greater than or equal to about 3.5,4.0, greater than or equal to about 4.5, greater than or equal to about5.5, greater than or equal to about 6.0, greater than or equal to about6.5, greater than or equal to about 7.0, greater than or equal to about7.5, greater than or equal to about 8.0, greater than or equal to about8.5, greater than or equal to about 9.0, greater than or equal to about9.5, or greater than or equal to about 10.0, greater than or equal toabout 10.5, greater than or equal to about 11.0, greater than or equalto about 11.5, greater than or equal to about 12.0, greater than orequal to about 12.5, greater than or equal to about 13.0, greater thanor equal to about 13.5, greater than or equal to about 14.0, greaterthan or equal to about 14.5, or greater than or equal to about 15.0. ThepI of a peptide can be within a range from about 3.0 to about 10.0,within a range from about 3.0 to about 6.0, or within a range from about4.0 to about 9.0.

In some embodiments, the pI (the pH at which the net charge of thepeptide is zero) of the peptides of this disclosure can be calculated bythe EMBOSS method. The pI value is the isoelectric point of fullyreduced form of protein sequences. The value can be calculated with theHenderson-Hasselbalch equation using EMBOSS scripts and a pKa tableprovided by the European Bioinformatics Institute. The EMBOSS method ofcalculating pI has been described by Rice et al. (EMBOSS: the EuropeanMolecular Biology Open Software Suite. Trends Genet. June 2000;16(6):276-7) and Carver et al. (The design of Jemboss: a graphical userinterface to EMBOSS. Bioinformatics. Sep. 22, 2003; 19(14):1837-43). Insome embodiments, peptides of the present disclosure with a pI valuegreater than 9 can have higher accumulation in the kidneys.

In some embodiments, the pI of the peptide influences its localizationwithin the kidney. For example, in certain embodiments, higher pI values(e.g., greater than or equal to about 7.5) promote localization and/orbinding to the glomerulus, while lower pI values (e.g., lower than 7.5)promote localization and/or binding to the proximal tubule. Accordingly,different localization patterns within the kidney can be achieved byvarying the pI of the peptide. In certain embodiments, the osmoticconcentration of the urine and/or urine flow rates have an impact onintratubular localization.

As another example, in various embodiments, the peptides of the presentdisclosure exhibit a charge distribution at neutral pH favorable forrenal localization, binding, and/or internalization. In certainembodiments, the peptide exhibits a substantially uniform chargedistribution. In alternative embodiments, the peptide exhibits anon-uniform charge distribution, e.g., including one or more regions ofconcentrated positive charge and/or one or more regions of concentratednegative charge. The charge distribution can impact the localization,binding and/or internalization of the peptide. For example, theglomerular capillary wall and/or slit processes are negatively charged,which in certain embodiments influences glomerular localization ofmiddle sized positively charged molecules (e.g., having a mass-averagemolecular weight (Mw) within a range from about 30 kDa to about 60 kDa),while being less likely to influence localization of smaller molecules(e.g., having a Mw less than 30 kDa) such as the peptides of the presentdisclosure. In certain embodiments, the charge distribution of thepeptide influences electrostatic interactions with a target, e.g., themegalin/cubulin receptor.

In yet another example, in various embodiments, the peptides of thepresent disclosure exhibit a molecular weight favorable for renaltargeting, localization, binding, accumulation, and/or internalization.In certain embodiments, the peptide comprises a mass-average molecularweight (Mw) less than or equal to about 1 kDa, less than or equal toabout 2 kDa, less than or equal to about 3 kDa, less than or equal toabout 4 kDa, less than or equal to about 5 kDa, less than or equal toabout 6 kDaor less than or equal to about 10 kDa, less than or equal toabout 20 kDa, less than or equal to about 30 kDa, less than or equal toabout 40 kDa, less than or equal to about 50 kDa, less than or equal toabout 60 kDa, or less than or equal to about 70 kDa. In certainembodiments, the peptide comprises a Mw within a range from about 0.5kDa to about 50 kDa, or within a range from about 0.5 kDa to about 60kDa.

In some embodiments, molecules (e.g., proteins or peptides) havingrelatively low Mw (e.g., less than or equal to about 1 kDa, less than orequal to about 2 kDa, less than or equal to about 3 kDa, less than orequal to about 4 kDa, less than or equal to about 5 kDa, less than orequal to about 10 kDa, less than or equal to about 20 kDa, less than orequal to about 30 kDa, or less than or equal to about 60 kDa) arerapidly targeted to, localized, bound, accumulated, and/or internalizedby the kidney. In certain embodiments, low Mw molecules are freelyfiltered, presented to the proximal tubules of the kidney, andoptionally taken up by megalin/cubulin receptors. In certainembodiments, low molecular weight molecules undergo endocyticreabsorption via the megalin/cubulin pathway and are then trafficked torenal tubular lysosomes for processing. In some embodiments, molecules(e.g., proteins or peptides) having higher Mw (e.g., greater than about70 kDa) are generally excluded from glomerular filtration, but can stillbe able to achieve interstitial localization via the microcirculation.

In a further example, in various embodiments, the peptides of thepresent disclosure exhibit stability at pH values favorable for renallocalization, binding, and/or internalization. A peptide can beconsidered to be stable at a certain pH if it is capable of performingits functional or therapeutic effect, is soluble, is resistant toprotease degradation, is resistant to reduction, retains secondary ortertiary structure, or a combination thereof. In certain embodiments,the peptide is stable at pH values less than or equal to about 3.0, lessthan or equal to about 3.5, 4.0, less than or equal to about 4.5, lessthan or equal to about 5.5, less than or equal to about 6.0, less thanor equal to about 6.5, less than or equal to about 7.0, less than orequal to about 7.5, less than or equal to about 8.0, less than or equalto about 8.5, less than or equal to about 9.0, less than or equal toabout 9.5, or less than or equal to about 10.0. In certain embodiments,the peptide is stable at pH values greater than or equal to about 3.0,greater than or equal to about 3.5, 4.0, greater than or equal to about4.5, greater than or equal to about 5.5, greater than or equal to about6.0, greater than or equal to about 6.5, greater than or equal to about7.0, greater than or equal to about 7.5, greater than or equal to about8.0, greater than or equal to about 8.5, greater than or equal to about9.0, greater than or equal to about 9.5, or greater than or equal toabout 10.0. In certain embodiments, the peptide is stable at pH valueswithin a range from about 3.0 to about 5.0, and/or within a range fromabout 5.0 to about 7.0.

As previously discussed, in some embodiments, the disulfide knotstructure of cystine-dense peptides confers improved stability over awide range of pH values, which can be advantageous for renalapplications. For example, stability at low pH values can beadvantageous in order to avoid cast formation leading to intratubularobstruction. In some embodiments, cast formation occurs viaco-precipitation of proteins with an endogenously produced glycoproteinknown as Tamm Horsall protein. In certain embodiments, thisprecipitation is affected by urinary pH and osmolality, as precipitationtypically occurs under acidic conditions (e.g., pH less than about 5)and high salt concentrations and/or osmolality. Alternatively or incombination, stability at low pH value can reduce or prevent lysosomaldegradation, which can improve delivery precision and avoid broadercellular or systemic toxicity.

Chemical Modifications

A peptide can be chemically modified one or more of a variety of ways.For example, N-methylation is one example of methylation that can occurin a peptide of the disclosure. In some embodiments, the peptide can bemutated to add function, delete function, or modify the in vivobehavior. One or more loops between the disulfide linkages can bemodified or replaced to include active elements from other peptides(such as described in Moore and Cochran, Methods in Enzymology, 503,p.223-251, 2012). Amino acids can also be mutated, such as to increasehalf-life or bioavailability, modify, add or delete binding behavior invivo, add new targeting function, modify surface charge andhydrophobicity, or allow conjugation sites. N-methylation is one exampleof methylation that can occur in a peptide of the disclosure. In someembodiments, the peptide can be modified by methylation on free amines.For example, full methylation can be accomplished through the use ofreductive methylation with formaldehyde and sodium cyanoborohydride.

A chemical modification can, for instance, extend the terminalhalf-life, the absorption half-life, the distribution half-life of apeptide, or change the biodistribution or pharmacokinetic profile. Achemical modification can comprise a polymer, a polyether, polyethyleneglycol, a biopolymer, a polyamino acid, a fatty acid, a dendrimer, an Fcregion, a simple saturated carbon chain such as palmitate ormyristolate, sugars, hyaluronic acid, or albumin. The chemicalmodification of a peptide with an Fc region can be a fusion Fc-peptide.A polyamino acid can include, for example, a polyamino acid sequencewith repeated single amino acids (e.g., polyglycine), and a polyaminoacid sequence with mixed polyamino acid sequences (e.g., gly-ala-gly-ala(SEQ ID NO: 634)) that can or cannot follow a pattern, or anycombination of the foregoing.

In some embodiments, the peptides of the present disclosure may bemodified such that the modification increases the stability and/or thehalf-life of the peptides. In some embodiments, the attachment of ahydrophobic moiety, such as to the N-terminus, the C-terminus, or aninternal amino acid, can be used to extend half-life of a peptide of thepresent disclosure. In other embodiments, the peptide of the presentdisclosure can include post-translational modifications (e.g.,methylation and/or amidation), which can affect, e.g., serum half-life.In some embodiments, simple carbon chains (e.g., by myristoylationand/or palmitylation) can be conjugated to the peptides. In someembodiments, for example, the simple carbon chains may render conjugatedpeptides easily separable from unconjugated material. For example,methods that may be used to separate the desired peptides of theinvention from unconjugated material include, but are not limited to,solvent extraction and reverse phase chromatography. In someembodiments, lipophilic moieties can be conjugated to the peptide andcan extend half-life through reversible binding to serum albumin.Moreover, the conjugated moieties can, e.g., be lipophilic moieties thatextend half-life of the peptides through reversible binding to serumalbumin. In some embodiments, the lipophilic moiety can be cholesterolor a cholesterol derivative including cholestenes, cholestanes,cholestadienes and oxysterols. In some embodiments, the peptides can beconjugated to myristic acid (tetradecanoic acid) or a derivativethereof. In other embodiments, the peptides of the present disclosureare coupled (e.g., conjugated) to a half-life modifying agent. Examplesof half-life modifying agents include but are not limited to: a polymer,a polyethylene glycol (PEG), a hydroxyethyl starch, polyvinyl alcohol, awater soluble polymer, a zwitterionic water soluble polymer, a watersoluble poly(amino acid), a water soluble polymer of proline, alanineand serine, a water soluble polymer containing glycine, glutamic acid,and serine, an Fc region, a fatty acid, palmitic acid, antibodies, or amolecule that binds to albumin.

In some embodiments, the first two N-terminal amino acids (GS or GG) ofSEQ ID NO: 1-SEQ ID NO: 41, SEQ ID NO: 471-SEQ ID NO: 529, SEQ ID NO:42-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ IDNO: 216-SEQ ID NO: 235, or SEQ ID NO: 530-SEQ ID NO: 549 can serve as aspacer or linker in order to facilitate conjugation or fusion to anothermolecule, as well as to facilitate cleavage of the peptide from suchconjugated or fused molecules. In some embodiments, the peptides of thepresent disclosure can be conjugated to other moieties that can modifyor effect changes to the properties of the peptides.

Active Agent Conjugates

Peptides according to the present disclosure can be conjugated or fusedto a peptide biological agent or other agent comprising amino acids(e.g., an antibody or antibody fragment, receptor or receptor fragment,ligand or ligand fragment, hormone or hormone fragment, growth factorsand growth factor fragments, biological toxins and fragments thereof, orother active portion of a peptide), a protein, a peptide, or to a smallmolecule, RNA, DNA, or other active agent molecular structure for use inthe treatment of renal diseases, disorders, or injuries. A peptideactive agent conjugate can be a peptide conjugated to an active agent byany mechanism described herein. For example, a peptide can be covalentlyconjugated to an active agent to form a peptide active agent conjugate.A peptide can be chemically conjugated to an active agent to form apeptide active agent conjugate. A peptide and active agent can beexpressed as a fusion protein to form a peptide active agent conjugate.For example, an antibody or fragment thereof and a peptide can beexpressed as a fusion protein to form a peptide active agent conjugate.For example, in certain embodiments, a peptide as described herein canbe fused to another molecule, such as an active agent that provides afunctional capability. The active agent can function as a renaltherapeutic agent, a renal protective agent, or renal prophylacticagent. A peptide can be conjugated with an active agent throughexpression of a vector containing the sequence of the peptide with thesequence of the active agent. In various embodiments, the sequence ofthe peptide and the sequence of the active agent are expressed from thesame Open Reading Frame (ORF). In various embodiments, the sequence ofthe peptide and the sequence of the active agent can comprise acontiguous sequence. Various vectors and recombinant systems known inthe art can be employed to make such fusion peptides. The peptide andthe active agent can each retain similar functional capabilities in thefusion peptide compared with their functional capabilities whenexpressed separately.

Furthermore, for example, in certain embodiments, the peptides describedherein are attached to another molecule, such as an active agent thatprovides a functional capability. In some embodiments, 1, 2, 3, 4, 5, 6,7, 8, 9, or 10 active agents can be linked to a peptide. Multiple activeagents can be attached by methods such as conjugating to multiple lysineresidues and/or the N-terminus, or by linking the multiple active agentsto a scaffold, such as a polymer or dendrimer and then attaching thatagent-scaffold to the peptide (such as described in Yurkovetskiy, A. V.,Cancer Res 75(16): 3365-72 (2015)).

Described herein are active agents that can be conjugated to thepeptides of the present invention for use in kidney disorders. In someembodiments, certain compounds or drugs are appropriate for use inkidney disorders. In some embodiments, certain drug classes may bepreferred for specific treatment depending on the indication ordisorder. As described herein, it is understood that certain activeagents are described in a non-limiting exemplary manner for use intreatments of kidney indications. One or more of such active agents canbe conjugated to a peptide of the present invention alone or incombination with one or more detectable agents described herein. In someembodiments, active agents that can be conjugated to any peptide of thisdisclosure can be classified by mechanism. For example, active agentscan belong to the class of anti-inflammatory drugs, immunosuppressive(immune suppression) drugs, analgesics/pain relief drugs, cell depletingagents/apoptosis modifiers, and tissue normalization (disease modifying)drugs.

Anti-inflammatory active agents can include, but are not limited to,corticosteroids, glucocorticoids, nonsteroidal anti-inflammatory drugs(NSAIDs), biologics, and other small molecules. Examples ofcorticosteroid active agents that can be conjugated to any peptide ofthis disclosure for delivery to the kidneys include triamcinolone,dexamethasone, budesonide, and triamcinolone acetonide. Examples ofNSAID active agents that can be conjugated to any peptide of thisdisclosure for delivery to the kidneys include naproxen and ibuprofen.Other active agents can include acetylsalicylic acid and acetaminophen.NSAID active agents can be further classified into COX2 inhibitors. Anexample of a COX2 inhibitor active agent directed to a prostaglandinpathway that can be conjugated to any peptide of this disclosure fordelivery includes celecoxib. An example of a COX2 inhibitor active agentwith anti-leukotriene receptor antagonist that can be conjugated to anypeptide of this disclosure for delivery includes montelukast. An exampleof a COX2 inhibitor active agent that can be conjugated to any peptideof this disclosure for delivery to the kidneys includes iguratimod.Biologic active agents can be further classified into active agents thatare IL-1 family inhibitors, IL-17 or IL-23 pathway inhibitors, IL-6family inhibitors, interferon receptor inhibitors, tumor necrosis factor(TNF) inhibitors, RANK pathway inhibitors, B cell inhibitors, anti-IgEactive agents, and co-stimulation inhibitors. An example of an IL-1family inhibitor active agent that can be conjugated to any peptide ofthis disclosure for delivery includes anakinra. An example of anIL-17/IL-23 pathway inhibitor active agent that can be conjugated to anypeptide of this disclosure for delivery includes secukinumab. An exampleof an IL-6 family inhibitor active agent that can be conjugated to anypeptide of this disclosure for delivery to the kidneys includessirukumab. An example of an interferon receptor inhibitor active agentthat can be conjugated to any peptide of this disclosure for delivery tothe kidneys includes anifrolumab. An example of a TNF inhibitor activeagent that can be conjugated to any peptide of this disclosure fordelivery includes infliximab or etanercept. An example of a RANK pathwayinhibitor active agent that can be conjugated to any peptide of thisdisclosure for delivery includes denosumab. An example of a B cellinhibitor active agent that can be conjugated to any peptide of thisdisclosure for delivery to the kidneys includes rituximab. An example ofan anti-IgE active agent that can be conjugated to any peptide of thisdisclosure for delivery to the kidneys includes omalizumab. An exampleof a co-stimulation inhibitor active agent that can be conjugated to anypeptide of this disclosure for delivery includes abatacept.

Pain relief active agents can include, but are not limited toanalgesics, counter-irritants, and pain receptor blocking drugs.Analgesics can be further classified into non-narcotic agents andnarcotic agents. An example of a non-narcotic active agent that can beconjugated to any peptide of this disclosure for delivery includesacetaminophen. An example of a narcotic active agent that can beconjugated to any peptide of this disclosure for delivery to the kidneysincludes oxycodone. Counter-irritant active agents can be furtherclassified as natural products. An example of a counter-irritant activeagent that can be conjugated to any peptide of this disclosure fordelivery include capsaicin, piperine, mustard oil, eugenol, andcurcumin, and capsaicin-like molecules like resiniferatoxin (RTX). Painreceptor blocking active agents can be further classified as TRPV4inhibitors. An example of a TRPV4 inhibitor active agent that can beconjugated to any peptide of this disclosure for delivery includesGSK2193874.

Apoptosis modifier active agents can include, but are not limited to,biologics and small molecules. Biologic apoptosis modifier active agentscan be further classified as Fas/FasL inhibitors, TNF/TNFR inhibitors,TRAIL/TRAILR inhibitors, TWEAK/Fn14 inhibitors, IL-1 inhibitors, IL-1receptor antagonists, growth factors, and sclerostin inhibitors. Anexample of a TNF/TNFR inhibitor active agent that can be conjugated toany peptide of this disclosure for delivery includes infliximab. Anexample of a TRAIL/TRAILR inhibitor active agent that can be conjugatedto any peptide of this disclosure for delivery includes osteoprotegerin.An example of a TWEAK/Fn14 inhibitor active agent that can be conjugatedto any peptide of this disclosure for delivery to the kidneys includesBIIB023. An example of an IL-1 receptor antagonist that can beconjugated to any peptide of this disclosure for delivery includesanakinra. An example of a growth factor active agent that can beconjugated to any peptide of this disclosure for delivery includesIGF-1. An example of a growth factor active agent that can be conjugatedto any peptide of this disclosure for delivery to the kidneys includesEGF. An example of a sclerostin inhibitor active agent that can beconjugated to any peptide of this disclosure for delivery includesromosozumab. Small molecule apoptosis modifier active agents can befurther classified as caspase inhibitors, iNOS inhibitors, surfactants,and bisphosphonates. An example of a caspase inhibitor active agent thatcan be conjugated to any peptide of this disclosure for deliveryincludes ZVAD-fmk. An example of an iNOS inhibitor active agent that canbe conjugated to any peptide of this disclosure for delivery includeS-methylisothiourea. An example of a surfactant active agent that can beconjugated to any peptide of this disclosure for delivery to the kidneysinclude P188. Moreover, the known class of drugs calledsenotherapeutics, also referred to as senolytics or senolytic drugs orsenolytic compounds, refers to small molecules that can selectivelyinduce death of senescent cells and for example by directly orindirectly inducing apoptosis in senescent cells. In addition,senolytics may also act via non-apoptotic mechanisms of cell deathincluding by necroptis, autophagic cell death, pyroptis andcaspase-independent cell death (Journal of Cell Science 127; 2135-2144(2014)). Such drugs can attenuate age-related deterioration of tissuesor organs. Examples of drugs that can be conjugated to any peptide ofthis disclosure to induce apoptosis or induce cell death vianon-apoptotic mechanisms include quercetin, dasatinib, bortezomib,carfilzomib, and navitoclax amongst other compounds disclosed herein.Additional examples are metformin, rapamycin, ABT-263, ABT-737, mTORmodulators, dasatinib, molecules that interact with FOXO, such as FOXO4peptide (Everts, “Can we hit the snooze button” Chemical and EngineeringNews, 95(10), 30-35,2017, Molecules that perturb the FOXO4 interactionwith p53, such as a FOXO4 peptide (Cell. 169(1): 132-147 (2017)). Otherexamples include dietary flavonols, small interfering RNA, or arapamycin analog such as RAD001. A further example of an active agentthat can be linked to any peptide of this disclosure is dimethylfumarate, which can be used for psoriatic arthritis or kidney fibrosis.Additional active agents are described in the following references:Aging Cell. August 2015; 14(4):644-58. doi: 10.1111/ace1.12344. EpubApr. 22, 2015. Kirkland J L (2013b) Translating advances from the basicbiology of aging into clinical application. Exp. Gerontol. 48, 1-5,Kirkland J L, Tchkonia T (2014) Clinical strategies and animal modelsfor developing senolytic agents. Exp. Gerontol. Oct. 28, 2014. pii:S0531-5565(14)00291-5, Tchkonia T, Zhu Y, van Deursen J, Campisi J,Kirkland J L (2013) Cellular senescence and the senescent secretoryphenotype: therapeutic opportunities. J. Clin. Invest. 123, 966-972,WO2016118859, WO2016118859, Pharmgenomics Pers Med. 2015; 8: 23-33, Renet al. Sci Rep. Apr. 7, 2016; 6:23968, Swanson et al. Nat Rev Rheumatol.June 2009; 5(6): 317-324, Oh et al. PLoS One. 2012; 7(10): e45870, andAdebajo, Ade, Wolf-Henning Boehncke, Dafna D. Gladman, and P J. Mease.Psoriatic Arthritis and Psoriasis: Pathology and Clinical Aspects.,2016. Internet resource.

Tissue normalization (disease modifying) active agents can include, butare not limited to, biologics and small molecules. Biologic activeagents can be further classified as chemokines (e.g. for stem cellrecruitment) and growth factors. An example of a tissue normalizationchemokine active agent that can be conjugated to any peptide of thisdisclosure for delivery to the kidney includes MIP-3α. An example of atissue normalization growth factor active agent that can be conjugatedto any peptide of this disclosure for delivery includes BMP-2 and BMP-7.Small molecule active agents can be further classified as flavonoids,ACE inhibitors, and anti-proliferative active agents. An example of atissue normalization flavonoid active agent that can be conjugated toany peptide of this disclosure for delivery to the kidney includesicariin. An example of a tissue normalization ACE inhibitor active agentthat can be conjugated to any peptide of this disclosure for delivery tothe kidneys includes captopril. An example of a tissue normalizationanti-proliferative active agent that can be conjugated to any peptide ofthis disclosure for delivery includes methotrexate.

TABLES 5 and 6 describe active agents for treatment of a kidney disorderthat can be conjugated to any peptide of the present disclosure to formpeptide-drug conjugates.

TABLE 5 Active Agents for Treatment of Kidney Disorders Active AgentClass Active Agent IL-6 Receptor Modulators Tocilizumab IL-6 ReceptorModulators Sarilumab IL-6 Receptor Modulators ALX-0061 IL-6 ReceptorModulators Sirukumab IL-6 Receptor Modulators Clazakizumab IL-6 ReceptorModulators Olokizumab IL-6 Receptor Modulators MEDI5117 IL-17Antagonists Secukinumab IL-17 Antagonists Brodalumab IL-17 AntagonistsIxekizumab Antagonists of p40 Subunit of IL- Ustekinumab 12/IL-23Antagonists of p40 Subunit of IL- Briakinumab 12/IL-23 Antagonists ofp19 Subunit of IL-23 Tildrakizumab Antagonists of p19 Subunit of IL-23Guselkumab IL-23 Antagonists Soluble IL-23 (or cytokine-binding homologyregion of soluble IL-23) IL-1 Antagonists Canakinumab IL-1 AntagonistsRilonacept IL-1 Antagonists Gevokizumab IL-1 Antagonists LY2189102 IL-1Antagonists Lentiviral-mediated RNAi IL-12 Antagonists IL-1 ReceptorAntagonists Anakinra IL-1 Receptor Antagonists MEDI-8968 IL-1 ReceptorAntagonists AMG-108 IL-1 Receptor Kineret Interleukins/Pro-InflammatoryPro-inflammatory IL-1α or IL-1β Cytokines Interleukins IL-8 InterleukinsIL-15 Interleukins IL-18 Interleukins IL-4 Interleukins IL-10Interleukins IL-13 Interleukins IL-17 p38 Inhibitors VX-745 p38Inhibitors BIRB 796 p38 Inhibitors SCIO-469 p38 Inhibitors VX-702 p38Inhibitors Pamapimod p38 Inhibitors ARRY-797 Corticosteroids17-monopropionate Corticosteroids Desciclesonide CorticosteroidsFlunisolide Corticosteroids 22-hydroxy intermediate budesonidederivative Corticosteroids 6β-hydroxy budesonide derivativeCorticosteroids Δ6-budesonide derivative Corticosteroids 23-hydroxybudesonide derivative Corticosteroids 16α-butryloxyprednisolonebudesonide derivative Corticosteroids 16α-hydroxyprednisolone budesonidederivative Corticosteroid (Beclomethasone) QVAR inhalationCorticosteroid (Budesonide) pulmicort respules Corticosteroid FloventHFA 44 Corticosteroid (Mometasone) Asmanex HFA Corticosteroid(Mometasone) Budesonide symbicort Corticosteroid Tixocortol pivalateCorticosteroid Ciclesonide Glucocorticoids 21-nortriamcincoloneacetonide Glucocorticoids Δ6-triamcinolone Glucocorticoids 6b-hydroxytriamcinolone acetonide Glucocorticoids 21-carboxy triamcinoloneacetonide Glucocorticoids 6b-OH, 21-COOH triamcinolone acetonideGlucocorticoids 6α fluorocortisol Glucocorticoids 9α fluorocortisolGlucocorticoids Δ1-dehydro configuration in prednisolone Glucocorticoids16-methylene dexamethasone derivative Glucocorticoids 16α-methyldexamethasone derivative Glucocorticoids 16β-methyl betamethasonederivative Glucocorticoids/Mineralocorticoids CortisolGlucocorticoids/Mineralocorticoids Betamethasone GlucocorticoidFluticasone propionate Steroid (flunisolide) Aerobid Steroid(flunisolide) Aerobid-M Steroid (flunisolide) Aerospan Steroid(Flunisolide) Fluticasone Furoate Steroid (Fluticasone) Flovent HFA 110Steroid (Fluticasone) Flovent HFA 220 Steroid (Fluticasone) FloventDiskus 50 Steroid (Fluticasone) Asmanex Steroid Betamethasone acetateSteroid Betamethasone sodium phosphate Steroid Betamethasone valerateSteroid Beclomethasone dipropionate Local Anesthetic procainehydrochloride Local Anesthetic novacain Anesthetic bupivacainehydrochloride Anesthetic lidocaine hydrochloride Local Anestheticropivacaine hydrochloride Analgesics Morphine Analgesics FentanylQuinazolines Feitinib/Iressa Quinazolines Sorafenib/Nexavar QuinazolinesLapatinib ditosylate/Tykerb/Tyverb Quinazolines Sunitinib/SutentQuinazolines Bortezomib/Velcade/Cytomib QuinazolinesEverolimus/Temsirolimus Quinazolines Inhibitors of IAPS QuinazolinesActivators of caspase pathway Quinazolines Activators of AKT pathwayQuinazolines Propylpeptidase inhibitors Quinazolines Activators of p53Quinazolines Inhibitors of anti-apoptotic protein inhibitors ProlylHydroxylase (PHD) Dimethyloxalylglycine (DMOG) Inhibitors ProlylHydroxylase (PHD) L-mimosine (L-mim) Inhibitors Aptamers Peptideaptamers Aptamers RNA aptamer A-p50 Aptamers Peptide A aptamer TrxLef1DAptamers Aptamer E07 Aptamers Aptamer gemcitabine polymers Aptamers RAGEAptamers Pegaptanib Proteosome Inhibitors Bortezomib ProteosomeInhibitors Carfilzomib Second Generation Proteosome Ixazomib InhibitorsSecond Generation Proteosome Delanzomib Inhibitors Second GenerationProteosome Oprozomib Inhibitors Second Generation Proteosome MarizomibInhibitors Apoptosis Inhibitors FLIP agonist Apoptosis Inhibitors nitricoxide synthase inhibitors Apoptosis Inhibitors caspase-3 inhibitors(Z-DEVD-fmk (SEQ ID NO: 635)) Apoptosis Inhibitors caspase-9 inhibitors(Z-LEHD-fmk (SEQ ID NO: 636)) Apoptosis Inhibitors Sclerostinantagonists Apoptosis Inhibitors/Growth Factor IGF-1 BCL-2 AgonistApoptosis Inhibitors Oblimersen BCL-2 Agonist Apoptosis Inhibitors GX01series of compounds BCL-2 Agonist Apoptosis Inhibitors BCL-2 smallmolecule antagonists BCL-2 Agonist Apoptosis Inhibitors Tetraocarcin-Aderivatives BCL-2 Agonist Apoptosis Inhibitors Chelerythrine BCL-2Agonist Apoptosis Inhibitors Antimycin A derivatives BCL-2 AgonistApoptosis Inhibitors HA14-1 BCL-2 Agonist Apoptosis Inhibitors Syntheticcompound antagonist of BH3 BCL-2 Agonist Apoptosis Inhibitors GenasenseBCL-2 Agonist Apoptosis Inhibitors ISIS 22783 BCL-2/BCL-XL AgonistApoptosis Bispecific Antisense Inhibitors Proapoptotic BCL-2 TargetingBax, Bak, Bid, Bad-derived BH3 Peptides Drugs Proapoptotic BCL-2Targeting SAHBs Drugs Proapoptotic BCL-2 Targeting BH3Is DrugsBCL-2/BCL-XL Agonist Apoptosis ABT-737 Inhibitors BCL-X InhibitorsApoptosis Modifiers Caspase-1 Inhibitors Apoptosis Modifiers Caspase-8Inhibitors Pan-caspase Caspase Inhibitor IDN-6556 Pan-caspase CaspaseInhibitor IDN-6734 Pan-caspase Caspase Inhibitor VX-799 Pan-caspaseInhibitor MX1013 Pan-caspase Caspase Inhibitor M-920 Pan-caspase CaspaseActivator MX-2060 derivatives Pan-caspase Caspase ActivatorsSmall-molecule compounds Pan-caspase Caspase Activators RGD peptidesPan-caspase inhibitors ZVAD-fmk Caspase-1 ICE Inhibitors IDN-11104Caspase-1 ICE Inhibitors VX-756 Caspase-3 Inhibitors M-826 Caspase-3Inhibitors M-791 Caspase-3 Inhibitors Immunocasp-3 Caspase-3 InhibitorsAd-G/iCasp3 Caspase-3 Inhibitors PEF-F8-CP3 Caspase-6 InhibitorsImmunocasp-6 Caspase-9 Inhibitors FKBP12/caspase-9 fusion protein IAPAntagonists BIR3 antagonists XIAP Antagonists Capped tripeptide XIAPAntagonists XIAP Antagonists Smac-mimetic compounds XIAP AntagonistsAEG35156/GEM ®640 XIAP Inhibitors Embelin XIAP Inhibitors XIAP antisenseand RNA constructs XIAP/cIAP-1/cIAP-2 Inhibitors Small molecule SMACmimetics IAP/Caspase Inhibitors HIV-Tat/polyarginine-conjugated SMACpeptides BIR2/Caspase-3 Inhibitors TWX024 BIR2 Inhibitors Polyphenylureaderivatives Survivin Targeting Drugs LY2181308 Survivin Targeting DrugsAd-Survivin T34A Xanthine Oxidase Inhibitors Allopurinol XanthineOxidase Inhibitors Febuxostat Xanthine Oxidase Inhibitors ZyloprinGrowth Factor bFGF Growth Factor IGF Growth Factor TFG-beta GrowthFactor BMP-2 Growth Factor BMP-9 Growth Factor BMP-13 Growth FactorBMP-7 Growth Factor BMP-3 inhibitors Growth Factor TFG-β1 Growth FactorOP-1 Growth Factor PDGF Growth Factor PTH Growth Factor PTHrP GrowthFactor MIP-3α Growth Factor FGF Growth Factor FGF-2 Growth Factor FGF-18Growth Factor TGF-β3 Growth Factor VEGF Growth Factor Wnt proteinsGrowth Factor EGF Growth Factor GM-CSF Flavonoid Icariin FlavonoidQuercetin Tyrosine Kinase Inhibitor (Lck/Btk Dasatinib Inhibitor) TRPV4Activators G5K1016790A TRPV4 Activators 4alpha-PDD TRPV4 InhibitorsHC-067047 TRPV4 Inhibitors GSK2193874 NSAID Ampion NSAID PhenylbutazoneNSAID Naproxen lysozyme conjugate NSAID Acetal salicylic acidImmunosuppresive and antiviral Leflunomide Quinolones Hydroxychloroquine(Plaquenil) Uricosurics Sulfinpyrazone MSC Matrix Collagen (e.g.,collagen type I, III, V, VI, VII and XV) MSC Matrix Fibrin MSC MatrixPolylactatous Extracellular Matrix Targeting Glycosaminoglycans (bothsulphated and non- sulphated glycosaminoglycans), glycoproteins andpolysaccharides Surfactant P188 and other surfactants Vascular GrowthFactor Angiopoetin Molecules for Bone Marrow Niches Bone morphogeniticproteins Catecholamines Epinephrine Catecholamines NorepinephrineMolecules for Bone Marrow Niches Jagged1 Notch Ligand osteopontinHormone parathyoid hormone Hormone Calcitonin Molecules for Bone MarrowNiches steel factor Glycoprotein Hormone thrombopoetin Vascular GrowthFactor vascular cell adhesion molecule 1 Chemokine Molecules for BoneCXCL12 Marrow Niches B Cell Targeting Agents Rituximab B Cell TargetingAgents BLys B Cell Targeting Agents TACI JAK Targeting AgentsTofacitinib Calcineurin Inhibitors Voclosporin COX-2 InhibitorsIguratimod COX-2 Inhibitors Montelukast COX-2 Inhibitors Rofecoxib COX-2Inhibitors Valdecoxib Interferon Receptor Inhibitors Anifrolumab IFN-αInhibitors Sifalimumab Anti-IgE Agents Omalizumab iNOS InhibitorsS-methylisothiourea CD20 Antagonists/B Cell Inhibitors Ocrelizumab BAFFAntagonists/B Cell Inhibitors Belimumab TNF Superfamily BAFF and APRILAtacicept Antagonists/B cell Inhibitors TNF-α Antagonists ThalidomideTNF-α Antagonists Lenalidomide TNF-α Antagonists Pomalidomide TNF-αAntagonists Pentocifylline TNF-α Antagonists Bupropion TNF AntagonistsLentiviral-mediated RNAi TNF Agonists Recombinant TNF-α TRAIL ReceptorAgonists HGS-ETR1 TRAIL Receptor Agonists HGS-ETR2 TRAIL ReceptorAgonists HGS-TR2J TRAIL Receptor Agonists PRO1762 TRAIL ReceptorAgonists TRA-8 CD95/Fas Agonists CD95-Fc Marine Bioactive CompoundsTRAIL-Resistance Overcoming Marine Bioactive Compounds Marine BioactiveCompounds mazamine A Marine Bioactive Compounds marine-derivedchroomycins Marine Bioactive Compounds carotenoids Marine BioactiveCompounds Aplysin Marine Bioactive Compounds Aplidin Marine BioactiveCompounds Siphonaxanthin Marine Bioactive Compounds pectinotoxin-2Anti-Complement Drugs Eculizumab PAR-2 Modulators Pepducin P2pal-18miR-2013 Blockers Anti-sense oligonucleotides Nrf2 Activator Dimethylfumarate p53 Targeting Drugs INGN201 p53 Targeting Drugs SCH58500 p53Targeting Drugs ONYX-015 p53 Targeting Drugs C-terminal p53 peptides p53Targeting Drugs CDB3 p53 Targeting Drugs CP31398 p53 Targeting DrugsPrima-1 p53 Targeting Drugs HPV E6-binding peptide aptamers p53Targeting Drugs Nutlins p53 Targeting Drugs Chalcones p53 TargetingDrugs Small peptides p53 Targeting Drugs Pifithrin-α p53 TargetingDrugs/Apoptosis QP1-1002 Modifiers (T cells) Endothelin-1 TargetingDrugs Astrasentan Immune Modulators Laquinimod Slow-acting antirheumaticdrugs (SAARDs) Colcrys Hormones parathyroid hormone Hormones growthhormone 11-beta hydroxysteroid dehydrogenases mineralocorticoidproopiomelanocortin fludrocortisonesoxycorticosterone acetate vaccinesfrom live attenuated viruses Aspirin Insulin Isonizaid Oral hypoglycemicagents Antacids carbamazepine cholestyramine colestipol ephedrineerythromyin mitotane oral contraceptives phenobarbital phenytoinrifampin troleandomycin Non-selective caspase inhibitor okadaic acidCamptothetic Staurosporine HFA Alvesco inhalation Breo Ellipta AdvairReactive Oxygen Species Targeting Drugs Cytokines/Growth FactorsTGF-beta NOD-like receptor protein 3- dependent caspase 1 TargetingDrugs NSAID Etoricoxib Apoptosis Modifiers MCL1 inhibitors TeriparatideBH3 mimetics AZD 4320 Carrier Proteins Low molecular weight human serumalbumin Ceramide Targeting Drugs Chondrogenic factors Anti-oxidativefactors A(1)AR agonist S1P(2)R antagonist Antimalarials BAX/BAKactivating drugs Selective GR Activators (SEGRAs) Rap1 Targeted DrugsCaspase-1 ICE Inhibitors VX-740 (Pralnacasan) Cathepsin K TargetingAgents Odanacatib TNF-α Antagonists CDP571 TNF-α Antagonists ISIS 104838Anti-Pain Drugs Duloxetine Cytokines/Growth Factors TGF-betaImmunosuppressants Rapamycin HIF-1α Modulators HIF-2α ModulatorsAngiotensin receptor blockers Angiotensin receptor blocker losartan(Cozaar) Hormones Adrenocorticotropic hormone Hormonescorticotropin-releasing hormone digitalis glycosides potassium-depletingdiuretics Coumarine anticoagulants NLRP3 Inflammosome Targeted MCC950Drugs NLRP3 Inflammosome Targeted BHB Drugs NLRP3 Inflammosome TargetedType I interferon Drugs NLRP3 Inflammosome Targeted IFN-beta Drugs NLRP3Inflammosome Targeted resveratrol Drugs NLRP3 Inflammosome Targetedarglabin Drugs NLRP3 Inflammosome Targeted CB2R agonist Drugs NLRP3Inflammosome Targeted MicroRNA-223 Drugs Immunosuppresive and rapamycinantiproliferative Bcl-2/Bcl-xL antagonist ABT-737 Tyrosine kinaseinhibitor dasatinib Oxycodone Janus kinase inhibitor Tofacitinib(generic name of Xeljanz)

TABLE 6 Further Active Agents for Treatment of Kidney Disorders ActiveAgent Class Active Agent Biguanide metformin Immunosuppressive mTORmodulators Immunosuppressive and FOXO4 peptide antiproliferativeAnti-inflammatory, Triptolide immunosuppressive Antioxidant Alpha-lipoicacid Checkpoint inhibitors Nivolumab Checkpoint inhibitors PembrolizumabCheckpoint inhibitors Pidilizumab Checkpoint inhibitors Bmx-936559Checkpoint inhibitors Atezolizumab Checkpoint inhibitors AvelumabAntibiotics Penicillins Penicillins Amoxicillin AntibioticsCephalosporins Cephalosporins Cephalexin Antibiotics MacrolidesMacrolides Azithromycin Antibiotics Fluoroquinolones FluoroquinolonesCiprofloxacin Antibiotics Sulfonamides Sulfonamides Co-trimoxazoleAntibiotics Tetracyclines Tetracyclines Doxycycline AntibioticsAminoglycosides Diuretics Loop Diuretics Diuretics Potassium SparingDiuretics Diuretics Chlorothiazide Diuretics Chlorthalidone DiureticsMetolazone Diuretics Indapamide Mineralocorticoid Renin Inhibitors ReninInhibitors aliskiren Renin Inhibitors pepstatin Renin Inhibitors statineRenin Inhibitors cgp2928 Renin Inhibitors remikiren Renin Inhibitorsenalkiren Renin Inhibitors zankiren SGLT modulator Dapagliflozin SGLTmodulator Canagliflozin SGLT modulator Empagliflozin Acetylsalicylicacid Steroid Beclomethasone monopropionate IL-17 inhibitor CaspaicinDeferasirox Olmesartan L-glutamic acid polymer Tirilazad Dietaryflavonols siRNA Rapamycin analogs RAD001 Counter-irritants PiperineCounter-irritants Mustard Oil Counter-irritants EugenolCounter-irritants Curcumin Counter-irritant capsaicin-likeResiniferatoxin (RTX) molecule

Further examples of active agents include but are not limited to: apeptide, an oligopeptide, a polypeptide, a peptidomimetic, apolynucleotide, a polyribonucleotide, a DNA, a cDNA, a ssDNA, a RNA, adsRNA, a micro RNA, an RNAi, an oligonucleotide, an antibody, a singlechain variable fragment (scFv or a single chain Fv), an antibodyfragment, an aptamer, a cytokine, an interferon, a hormone, an enzyme, agrowth factor, alpha-lipoic acid (to prevent nephrotoxicity to tubularcells after chemotherapy (e.g. cisplatin) or administration of an NSAID(e.g. indomethacin), a checkpoint inhibitor, nivolumab, pembrolizumab,pidilizumab, bmx-936559, atezolizumab, avelumab, a PD-1 inhibitor, aPD-L1 inhibitor, a CTLA4 inhibitor, a CD antigen, aa chemokine, aneurotransmitter, an ion channel inhibitor, a G-protein coupled receptorinhibitor, a G-protein coupled receptor activator, a chemical agent, aradiosensitizer, a radioprotectant, a radionuclide, a therapeutic smallmolecule, a steroid, a corticosteroid, an anti-inflammatory agent, animmune modulator, a complement fixing peptide or protein, a tumornecrosis factor inhibitor, a tumor necrosis factor activator, a tumornecrosis factor receptor family agonist, a tumor necrosis receptorantagonist, a tumor necrosis factor (TNF) soluble receptor or antibody,caspase protease activator or inhibitor, an NF-κB a RIPK1 and/or RIPK3inhibitor or activator (e.g., through Toll-like receptors (TLRs) TLR-3and/or TLR-4, or T-cell receptor (TCR) and the like), a death-receptorligand (e.g., Fas ligand) activator or inhibitor, TNF receptor family(e.g., TNFR1, TNFR2, lymphotoxin β receptor/TNFRS3, OX40/TNFRSF4,CD40/TNFRSF5, Fas/TNFRSF6, decoy receptor 3/TNFRSF6B, CD27/TNFRSF7,CD30/TNFRSF8, 4-1BB/TNFRSF9, DR4 (death receptor 4/TNFRS10A), DR5 (deathreceptor 5/TNFRSF10B), decoy receptor 1/TNFRSF10C, decoy receptor2/TNFRSF10D, RANK (receptor activator of NF-kappa B/TNFRSF11A), OPG(osteoprotegerin/TNFRSF11B), DR3 (death receptor 3/TNFRSF25), TWEAKreceptor/TNFRSF12A, TAC1/TNFRSF13B, BAFF-R (BAFF receptor/TNFRSF13C),HVEM (herpes virus entry mediator/TNFRSF14), nerve growth factorreceptor/TNFRSF16, BCMA (B cell maturation antigen/TNFRSF17), GITR(glucocorticoid-induced TNF receptor/TNFRSF18), TAJ (toxicity and JNKinducer/TNFRSF19), RELT/TNFRSF19L, DR6 (death receptor 6/TNFRSF21),TNFRSF22, TNFRSF23, ectodysplasin A2 isoform receptor/TNFRS27,ectodysplasin 1, and anhidrotic receptor, a TNF receptor superfamilyligand including—TNF alpha, lymphotoxin-α, tumor necrosis factormembrane form, tumor necrosis factor shed form, LIGHT, lymphotoxin β₂α₁heterotrimer, OX-40 ligand, compound 1 [PMID: 24930776], CD40 ligand,Fas ligand, TL1A, CD70, CD30 ligand, TRAF1, TRAF2, TRAF3, TRAIL, RANKligand, APRIL, BAFF, B and T lymphocyte attenuator, NGF, BDNF,neurotrophin-3, neurotrophin-4, TL6, ectodysplasin A2, ectodysplasinA1—a TIMP-3 inhibitor, a BCL-2 family inhibitor, navitoclax (Aging Cell.15(3): 428-435. (2016)), an IAP disruptor, a protease inhibitor, anamino sugar, a chemotherapeutic (whether acting through an apoptotic ornon-apoptotic pathway) (Ricci et al. Oncologist 11(4):342-57 (2006)), acytotoxic chemical, a toxin, a tyrosine kinase inhibitor (e.g. imatinibmesylate), protons, bevacuzimab (antivascular agent), erlotinib (EGFRinhibitor), QPI-1002, QM56, SVT016426 (QM31), 16/86 (third generationferrostatin), BASP siRNA, CCX140, BIIB023, CXA-10, alkaline phosphatase,Dnmt1 inhibitor, THR-184, lithium, formoterol, IL-22, EPO and EPOderivatives, agents that stimulate erthyropoietin such as epoeitn alfaor darbepoietin alfa, PDGF inhibitors, CRMD-001, Atrasentan, Tolvaptan,RWJ-676070, Abatacept, Sotatercept, the binding site of theextracellular domain of the activing receptor 2A, an anti-infectiveagent, an antibiotic; such as gentamicin, vancomycin, minocin ormitomyclin, penicillins (such as amoxicillin), cephalosporins (such ascephalexin), macrolides (such as azithromycin), fluoroquinolones (suchas ciprofloxacin), sulfonamides (such as co-trimoxazole), tetracyclines(such as doxycycline), aminoglycosides, an anti-infective agent, anantibiotic, an anti-viral agent, an anti-fungal agent, anaminoglycoside, a nonsteroidal anti-inflammatory drug (NSAID), a statin,a nanoparticle, a liposome, such as ketorolac or ibuprofen, animmunosuppresant such tacrolimus, mycophenolic acid (e.g., mycophenolatemofetil), cyclosporine A, or azathioprine, a diuretic drug includingthiazides, loop diuretics, and potassium sparing diuretics, bumetanide,ethacrynic acid, furosemide, torsemide, glucose, mannitol, amiloride,spironolactone, eplerenone, triamterene, potassium canrenoate,bendroflumethiazide, chlorothiazide, chlorthalidone, metolazone,indapamide, hydrochlorothiazide, vasopressin, amphotericin B,acetazolamide, tovaptan, conivaptan, dopamine, dorzolamide,bendrolumethiazide, hydrochlorothiazide, caffeine, theophylline, ortheobromine, a statin, a senolytic such as navitoclax or obatoclax, acorticosteroid such as prednisone, betamethasone, fludrocortisone,deoxycorticosterone, aldosterone, cortisone, hydrocortisone,belcometasone, dexamethasone, mometasone, fluticasone, prednisolone,methylprednisolone, triamcinolone acetonide or triamcinolone, aglucocorticoid, a mineralocorticoid, such as aldosterone andflucrocortisone, a liposome, renin, renin inhibitors such as aliskiren,pepstatin, statine, cgp2928, remikiren, enalkiren, zankiren,angiotensin, ACE inhibitors such as ramipril, captopril, lisinopril,benazepril, quinapril, fosinopril, trandolapril, moexipril, enalaprilat,enalapril maleate, or perindopril erbumine, mediator of apoptosis,mediator of fibrosis, drug that targets p53, Apaf-1 inhibitor, RIPK1inhibitor, RIPK3 inhibitor, inhibitor of IL17, inhibitor of IL6,inhibitor of IL23, inhibitor of CCR2, nitrated fatty acids, angiotensinblockers, agonists of the ALK3 receptor, retinoic acid, SGLT2 modulator,such as Dapagliflozin, canagliflozin, and empagliflozin, a polymer, abiopolymer, a polysaccharide, a proteoglycan, a glycosaminoglycan,polyethylene glycol, a lipid, a dendrimer, a fatty acid, or an Fc domainor an Fc region, or an active fragment or a modification thereof. Anycombination of the above active agents can be co-delivered with peptidesor peptide conjugates of this disclosure. Additionally, in someembodiments, other co-therapies such as proton therapy or ablativeradiotherapy can be administered to a subject in need thereof along withpeptides or peptide conjugates of this disclosure. In some embodiments,the peptide is covalently or non-covalently linked to an active agent,e.g., directly or via a linker. TNF blockers suppress the immune systemby blocking the activity of TNF, a substance in the body that can causeinflammation and lead to immune-system diseases, such as lupus, Crohn'sdisease, and ulcerative colitis. The drugs in this class includeRemicade (infliximab), Enbrel (etanercept), Humira (adalimumab), Cimzia(certolizumab pegol) and Simponi (golimumab). The peptide disclosedherein can be used to home, distribute to, target, directed to, isretained by, accumulate in, migrate to, and/or bind to kidneys, and thusalso be used for localizing the attached or fused active agent.Furthermore, chlorotoxin peptide can be internalized in cells(Wiranowska, M., Cancer Cell Int., 11: 27 (2011)). Therefore, cellularinternalization, subcellular localization, and intracellular traffickingafter internalization of the peptide itself, or an active agent peptideconjugate or fusion peptide can be important factors in the efficacy ofan active agent conjugate or fusion. (Ducry, L., Antibody DrugConjugates (2013); and Singh, S. K., Pharm Res. 32(11): 3541-3571(2015)). Exemplary linkers suitable for use with the embodiments hereinare discussed in further detail below.

The peptides or peptide-active agent fusions of the present disclosurecan also be conjugated to other moieties that can serve other roles,such as providing an affinity handle (e.g., biotin) for retrieval of thepeptides from tissues or fluids. For example, peptides or peptide-activeagent fusions of the present disclosure can also be conjugated tobiotin. In addition to extension of half-life, biotin could also act asan affinity handle for retrieval of peptides or peptide-active agentfusions from tissues or other locations. In some embodiments,fluorescent biotin conjugates that can act both as a detectable labeland an affinity handle can be used. Non limiting examples ofcommercially available fluorescent biotin conjugates include Atto425-Biotin, Atto 488-Biotin, Atto 520-Biotin, Atto-550 Biotin, Atto565-Biotin, Atto 590-Biotin, Atto 610-Biotin, Atto 620-Biotin, Atto655-Biotin, Atto 680-Biotin, Atto 700-Biotin, Atto 725-Biotin, Atto740-Biotin, fluorescein biotin, biotin-4-fluorescein,biotin-(5-fluorescein) conjugate, and biotin-B-phycoerythrin, Alexafluor 488 biocytin, Alexa flour 546, Alexa Fluor 549, lucifer yellowcadaverine biotin-X, Lucifer yellow biocytin, Oregon green 488 biocytin,biotin-rhodamine and tetramethylrhodamine biocytin. In some otherexamples, the conjugates could include chemiluminescent compounds,colloidal metals, luminescent compounds, enzymes, radioisotopes, andparamagnetic labels. In some embodiments, the peptide-active agentfusions described herein can be attached to another molecule. Forexample, the peptide sequence also can be attached to another activeagent (e.g., small molecule, peptide, polypeptide, polynucleotide,antibody, aptamer, cytokine, growth factor, neurotransmitter, an activefragment or modification of any of the preceding, fluorophore,radioisotope, radionuclide chelator, acyl adduct, chemical linker, orsugar, etc.). In some embodiments, the peptide can be fused with, orcovalently or non-covalently linked to an active agent.

Additionally, more than one peptide sequence can be present on or fusedwith a particular peptide. A peptide can be incorporated into abiomolecule by various techniques, for example by a chemicaltransformation, such as the formation of a covalent bond, such as anamide bond, or by solid phase or solution phase peptide synthesis, or bypreparing a nucleic acid sequence encoding the biomolecule, wherein thenucleic acid sequence includes a subsequence that encodes the peptide.The subsequence can be in addition to the sequence that encodes thebiomolecule, or can substitute for a subsequence of the sequence thatencodes the biomolecule.

In some embodiments, the peptides of the present disclosure are coupled(e.g., conjugated) to other moieties that, e.g., can modify or effectchanges to the properties of the peptides. For example, in certainembodiments, the peptides described herein are attached to anothermolecule, such as an active agent that provides a functional capability.Examples of active agents include but are not limited to: a peptide, anoligopeptide, a polypeptide, a polynucleotide, a polyribonucleotide, aDNA, a cDNA, a ssDNA, a RNA, a dsRNA, a micro RNA, an oligonucleotide,an antibody fragment, a single chain Fv, an aptamer, a cytokine, anenzyme, a growth factor, a chemokine, a neurotransmitter, a chemicalagent, a fluorophore, a metal, a metal chelate, an X-ray contrast agent,a PET agent, a radioisotope, a photosensitizer, a radiosensitizer, aradionuclide chelator, a therapeutic small molecule, a steroid, acorticosteroid, an anti-inflammatory agent, an immune modulator, aprotease inhibitor, an amino sugar, a chemotherapeutic, a cytotoxicchemical, a toxin, a tyrosine kinase inhibitor, an anti-infective agent,an antibiotic, an anti-viral agent, an anti-fungal agent, anaminoglycoside, a nonsteroidal anti-inflammatory drug (NSAID) such asketorolac or ibuprofen, a statin, a nanoparticle, a liposome, a polymer,a biopolymer, a polysaccharide, a proteoglycan, a glycosaminoglycan, adendrimer, a fatty acid, or an Fc region, or an active fragment or amodification thereof. In some embodiments, the peptide is covalently ornon-covalently linked to an active agent, e.g., directly or via alinker. Exemplary linkers suitable for use with the embodiments hereinare discussed in further detail below.

In some embodiments, the active agent interacts with a renal ionchannel, inhibits a protease, has antimicrobial activity, has anticanceractivity, has anti-inflammatory activity, induces ischemicpreconditioning or acquired cytoresistance, produces a protective ortherapeutic effect on a kidney of the subject, reduces a clearance rateof the composition, or a combination thereof. Optionally, the activeagent is a renal therapeutic agent, such as a renal protective agent orrenal prophylactic agent that induces ischemic preconditioning and/oracquired cytoresistance in a kidney of a subject. Additional detailsregarding renal therapeutic agents are provided below.

In some embodiments, the peptides of the present disclosure can bemodified such that the modification increases the stability and/or thehalf-life of the peptides. In some embodiments, the attachment of ahydrophobic moiety, such as to the N-terminus, the C-terminus, or on aninternal amino acid, can be used to extend half-life of a peptide of thepresent disclosure. In some embodiments, simple carbon chains (e.g., bymyristoylation and/or palmitylation) can be conjugated to the fusionproteins or peptides. In some embodiments, the simple carbon chains canrender the peptides easily separable from the unconjugated material. Forexample, methods that can be used to separate the peptides from theunconjugated material include, but are not limited to, solventextraction and reverse phase chromatography. The lipophilic moieties canextend half-life through reversible binding to serum albumin. Theconjugated moieties can, e.g., be lipophilic moieties that extendhalf-life of the peptides through reversible binding to serum albumin.In some embodiments, simple carbon chains (e.g., by myristoylation) canbe conjugated to the peptides. In some embodiments, the lipophilicmoiety can be cholesterol or a cholesterol derivative includingcholestenes, cholestanes, cholestadienes and oxysterols. In someembodiments, the peptides can be conjugated to myristic acid(tetradecanoic acid) or a derivative thereof.

Detectable Agent Conjugates

Described herein are agents that can be conjugated to the peptides ofthe present invention for use in detection and tracing either kidneydisorders, or both. As described herein, it is understood that certainactive agents are described in a non-limiting exemplary manner for usein diagnostics, aiding surgery and treatment, prognosis and tracking ofprogress or remission of kidney disorders, diseases or injury. One ormore of such detectable agents can be conjugated to a peptide of thepresent invention alone or in combination with one or more active agentsdescribed herein. Moreover some detectable agents (e.g., radionuclides,radioisotopes, radiosensitizers and photosensitizers amongst others) mayalso exert therapeutic activity as well. A peptide can be conjugated toan agent used in imaging, research, therapeutics, theranostics,pharmaceuticals, chemotherapy, chelation therapy, targeted drugdelivery, and radiotherapy. The agent can be a detectable agent. In someembodiments, a peptide of the present invention is conjugated todetectable agents, such as a metal, a radioisotope, a dye, fluorophore,or another suitable material that can be used in imaging. Non-limitingexamples of radioisotopes include alpha emitters, beta emitters,positron emitters, and gamma emitters. In some embodiments, the metal orradioisotope is selected from the group consisting of actinium,americium, bismuth, cadmium, cesium, cobalt, europium, gadolinium,iridium, lead, lutetium, manganese, palladium, polonium, radium,ruthenium, samarium, strontium, technetium, thallium, and yttrium. Insome embodiments, the metal is actinium, bismuth, lead, radium,strontium, samarium, or yttrium. In some embodiments, the radioisotopeis actinium-225 or lead-212. In some embodiments, the fluorophore is afluorescent agent emitting electromagnetic radiation at a wavelengthbetween 650 nm and 4000 nm, such emissions being used to detect suchagent. In some embodiments the fluorophore is a fluorescent agent isselected from the group consisting of non-limiting examples offluorescent dyes that could be used as a conjugating molecule (or asapplied to each class of molecules) in the present disclosure includeDyLight-680, DyLight-750, VivoTag-750, DyLight-800, IRDye-800,VivoTag-680, Cy5.5, ZQ800, or indocyanine green (ICG class of dyes). Insome embodiments, near infrared dyes often include cyanine dyes.Additional non-limiting examples of fluorescent dyes for use as aconjugating molecule in the present disclosure include acradine orangeor yellow, Alexa Fluors and any derivative thereof, 7-actinomycin D,8-anilinonaphthalene-1-sulfonic acid, ATTO dye and any derivativethereof, auramine-rhodamine stain and any derivative thereof,bensantrhone, bimane, 9-10-bis(phenylethynyl)anthracene,5,12-bis(phenylethynyl)naththacene, bisbenzimide, brainbow, calcein,carbodyfluorescein and any derivative thereof,1-chloro-9,10-bis(phenylethynyl)anthracene and any derivative thereof,DAPI, DiOC6, DyLight Fluors and any derivative thereof, epicocconone,ethidium bromide, FlAsH-EDT2, Fluo dye and any derivative thereof,FluoProbe and any derivative thereof, Fluorescein and any derivativethereof, Fura and any derivative thereof, GelGreen and any derivativethereof, GelRed and any derivative thereof, fluorescent proteins and anyderivative thereof, m isoform proteins and any derivative thereof suchas for example mCherry, hetamethine dye and any derivative thereof,hoeschst stain, iminocoumarin, indian yellow, indo-1 and any derivativethereof, laurdan, lucifer yellow and any derivative thereof, luciferinand any derivative thereof, luciferase and any derivative thereof,mercocyanine and any derivative thereof, nile dyes and any derivativethereof, perylene, phloxine, phyco dye and any derivative thereof,propium iodide, pyranine, rhodamine and any derivative thereof,ribogreen, RoGFP, rubrene, stilbene and any derivative thereof,sulforhodamine and any derivative thereof, SYBR and any derivativethereof, synapto-pHluorin, tetraphenyl butadiene, tetrasodium tris,Texas Red, Titan Yellow, TSQ, umbelliferone, violanthrone, yellowfluorescent protein and YOYO-1. Other Suitable fluorescent dyes include,but are not limited to, fluorescein and fluorescein dyes (e.g.,fluorescein isothiocyanine or FITC, naphthofluorescein,4′,5′-dichloro-2′,7′-dimethoxyfluorescein, 6-carboxyfluorescein or FAM,etc.), carbocyanine, merocyanine, styryl dyes, oxonol dyes,phycoerythrin, erythrosin, eosin, rhodamine dyes (e.g.,carboxytetramethyl-rhodamine or TAMRA, carboxyrhodamine 6G,carboxy-X-rhodamine (ROX), lissamine rhodamine B, rhodamine 6G,rhodamine Green, rhodamine Red, tetramethylrhodamine (TMR), etc.),coumarin and coumarin dyes (e.g., methoxycoumarin, dialkylaminocoumarin,hydroxycoumarin, aminomethylcoumarin (AMCA), etc.), Oregon Green Dyes(e.g., Oregon Green 488, Oregon Green 500, Oregon Green 514, etc.),Texas Red, Texas Red-X, SPECTRUM RED, SPECTRUM GREEN, cyanine dyes(e.g., CY-3, Cy-5, CY-3.5, CY-5.5, etc.), ALEXA FLUOR dyes (e.g., ALEXAFLUOR 350, ALEXA FLUOR 488, ALEXA FLUOR 532, ALEXA FLUOR 546, ALEXAFLUOR 568, ALEXA FLUOR 594, ALEXA FLUOR 633, ALEXA FLUOR 660, ALEXAFLUOR 680, etc.), BODIPY dyes (e.g., BODIPY FL, BODIPY R6G, BODIPY TMR,BODIPY TR, BODIPY 530/550, BODIPY 558/568, BODIPY 564/570, BODIPY576/589, BODIPY 581/591, BODIPY 630/650, BODIPY 650/665, etc.), IRDyes(e.g., IRD40, IRD 700, IRD 800, etc.), indocyanine green dyes and thelike. For each of the above listed fluorescent dyes various activatedforms can be used for conjugation and the like. Additional suitabledetectable agents are described in PCT/US14/56177. Non-limiting examplesof radioisotopes include alpha emitters, beta emitters, positronemitters, and gamma emitters. In some embodiments, the metal orradioisotope is selected from the group consisting of actinium,americium, bismuth, cadmium, cesium, cobalt, europium, gadolinium,iridium, lead, lutetium, manganese, palladium, polonium, radium,ruthenium, samarium, strontium, technetium, thallium, and yttrium. Insome embodiments, the metal is actinium, bismuth, lead, radium,strontium, samarium, or yttrium. In some embodiments, the radioisotopeis actinium-225 or lead-212.

Other embodiments of the present disclosure provide peptides conjugatedto a radiosensitizer or photosensitizer. Examples of radiosensitizersinclude but are not limited to: ABT-263, ABT-199, WEHI-539, paclitaxel,carboplatin, cisplatin, oxaliplatin, gemcitabine, etanidazole,misonidazole, tirapazamine, and nucleic acid base derivatives (e.g.,halogenated purines or pyrimidines, such as 5-fluorodeoxyuridine).Examples of photosensitizers include but are not limited to: fluorescentmolecules or beads that generate heat when illuminated, porphyrins andporphyrin derivatives (e.g., chlorins, bacteriochlorins,isobacteriochlorins, phthalocyanines, and naphthalocyanines),metalloporphyrins, metallophthalocyanines, angelicins,chalcogenapyrrillium dyes, chlorophylls, coumarins, flavins and relatedcompounds such as alloxazine and riboflavin, fullerenes, pheophorbides,pyropheophorbides, cyanines (e.g., merocyanine 540), pheophytins,sapphyrins, texaphyrins, purpurins, porphycenes, phenothiaziniums,methylene blue derivatives, naphthalimides, nile blue derivatives,quinones, perylenequinones (e.g., hypericins, hypocrellins, andcercosporins), psoralens, quinones, retinoids, rhodamines, thiophenes,verdins, xanthene dyes (e.g., eosins, erythrosins, rose bengals),dimeric and oligomeric forms of porphyrins, and prodrugs such as5-aminolevulinic acid. Advantageously, this approach allows for highlyspecific targeting of diseased cells (e.g., cancer cells) using both atherapeutic agent (e.g., drug) and electromagnetic energy (e.g.,radiation or light) concurrently. In some embodiments, the peptide iscovalently or non-covalently linked to the agent, e.g., directly or viaa linker. Exemplary linkers suitable for use with the embodiments hereinare discussed in further detail below.

Linkers

Peptides according to the present disclosure that home, target, migrateto, are retained by, accumulate in, and/or bind to, or are directed tothe kidney can be attached to another moiety (e.g., an active agent),such as a small molecule, a second peptide, a protein, an antibody, anantibody fragment, an aptamer, polypeptide, polynucleotide, afluorophore, a radioisotope, a radionuclide chelator, a polymer, abiopolymer, a fatty acid, an acyl adduct, a chemical linker, or sugar orother active agent described herein through a linker, or directly in theabsence of a linker.

A peptide can be directly attached to another molecule by a covalentattachment. For example, the peptide is attached to a terminus of theamino acid sequence of a larger polypeptide or peptide molecule, or isattached to a side chain, such as the side chain of a lysine, serine,threonine, cysteine, tyrosine, aspartic acid, a non-natural amino acidresidue, or glutamic acid residue. The attachment can be via an amidebond, an ester bond, an ether bond, a carbamate bond, a carbon-nitrogenbond, a triazole, a macrocycle, an oxime bond, a hydrazone bond, acarbon-carbon single double or triple bond, a disulfide bond, or athioether bond. In some embodiments, similar regions of the disclosedpeptide(s) itself (such as a terminus of the amino acid sequence, anamino acid side chain, such as the side chain of a lysine, serine,threonine, cysteine, tyrosine, aspartic acid, a non-natural amino acidresidue, or glutamic acid residue, via an amide bond, an ester bond, anether bond, a carbamate bond, a carbon-nitrogen bond, a triazole, amacrocycle, an oxime bond, a hydrazone bond, a carbon-carbon singledouble or triple bond, a disulfide bond, or a thioether bond, or linkeras described herein) can be used to link other molecules.

Attachment via a linker can involve incorporation of a linker moietybetween the other molecule and the peptide. The peptide and the othermolecule can both be covalently attached to the linker. The linker canbe cleavable, labile, non-cleavable, stable, self-immolating,hydrophilic, or hydrophobic. As used herein, the term “non-cleavable”(such as used in association with an amide, cyclic, or carbamate linkeror as otherwise as described herein) is often used by a skilled artisanto distinguish a relatively stable structure from one that is morelabile or “cleavable” (e.g., as used in association with cleavablelinkers that may be dissociated or cleaved structurally by enzymes,proteases, self-immolation, pH, reduction, hydrolysis, certainphysiologic conditions, or as otherwise described herein). It isunderstood that “non-cleavable” linkers offer stability against cleavageor other dissociation as compared to “cleavable” linkers, and the termis not intended to be considered an absolute non-cleavable ornon-dissociative structure under any conditions. Consequently, as usedherein, a “non-cleavable” linker is also referred to as a “stable”linker. The linker can have at least two functional groups with onebonded to the peptide, the other bonded to the other molecule, and alinking portion between the two functional groups.

Non-limiting examples of the functional groups for attachment caninclude functional groups capable of forming an amide bond, an esterbond, an ether bond, a carbonate bond, a carbamate bond, or a thioetherbond. Non-limiting examples of functional groups capable of forming suchbonds can include amino groups; carboxyl groups; hydroxyl groups;aldehyde groups; azide groups; alkyne and alkene groups; ketones;hydrazides; acid halides such as acid fluorides, chlorides, bromides,and iodides; acid anhydrides, including symmetrical, mixed, and cyclicanhydrides; carbonates; carbonyl functionalities bonded to leavinggroups such as cyano, succinimidyl, and N-hydroxysuccinimidyl; hydroxylgroups; sulfhydryl groups; and molecules possessing, for example, alkyl,alkenyl, alkynyl, allylic, or benzylic leaving groups, such as halides,mesylates, tosylates, triflates, epoxides, phosphate esters, sulfateesters, and besylates.

Non-limiting examples of the linking portion can include alkylene,alkenylene, alkynylene, polyether, such as polyethylene glycol (PEG),hydroxy carboxylic acids, oligoethylene glycol, polyester, polyamide,polyamino acids, polypeptides, cleavable peptides, valine-citrulline,aminobenzylcarbamates, D-amino acids, and polyamine, any of which beingunsubstituted or substituted with any number of substituents, such ashalogens, hydroxyl groups, sulfhydryl groups, amino groups, nitrogroups, nitroso groups, cyano groups, azido groups, sulfoxide groups,sulfone groups, sulfonamide groups, carboxyl groups, carboxaldehydegroups, imine groups, alkyl groups, halo-alkyl groups, alkenyl groups,halo-alkenyl groups, alkynyl groups, halo-alkynyl groups, alkoxy groups,aryl groups, aryloxy groups, aralkyl groups, arylalkoxy groups,heterocyclyl groups, acyl groups, acyloxy groups, carbamate groups,amide groups, urethane groups, epoxides, and ester groups.

A peptide and drug conjugated via a linker is described with the formulaPeptide-A-B-C-Drug, wherein the linker is A-B-C. A can be stable amidelink is an amine on the peptide and the linker and can be achieved via atetrafluorophenyl (TFP) ester or an NHS ester. B can be (—CH2—)_(x)— ora short PEG (—CH₂CH₂O—)_(x) (x is 1-10), and C can be the ester bond tothe hydroxyl or carboxylic acid on the drug. In some embodiments, C canrefer to the “cleavable” or “stable” part of the linker. In otherembodiments, A can also be the “cleavable” part. In some embodiments, Acan be amide, carbamate, thioether via maleimide or bromoacetamide,triazole, oxime, or oxacarboline. The cleaved active agent or drug canretain the chemical structure of the active agent before cleavage, orcan be modified as a result of cleavage. Moreover, depending on thedesired therapeutic properties of the peptide-drug conjugate, suchactive agent can be active while linked to the peptide, remain activeafter cleavage or become inactivated, be inactive while linked to thepeptide, or it can be activated upon cleavage.

In some embodiments, peptide conjugates have stable linkers. A peptideof the disclosure can be expressed recombinantly or chemicallysynthesized. The peptide can be conjugated to a detectable agent or anactive agent via a stable linker, such as an amide linkage or acarbamate linkage. The peptide can be conjugated to a detectable agentor an active agent via a stable linker, such as an amide bond usingstandard 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) ordicylcohexylcarbodiimide (DCC) based chemistry or thionyl chloride orphosphorous chloride-based bioconjugation chemistries. A stable linkermay or may not be cleaved in buffer over extended periods of time (e.g.,hours, days, or weeks). A stable linker may or may not be cleaved inbody fluids such as plasma or synovial fluid over extended periods oftime (e.g., hours, days, or weeks). A stable linker, may or may not becleaved after exposure to enzymes, reactive oxygen species, otherchemicals or enzymes that can be present in cells (e.g., macrophages),cellular compartments (e.g., endosomes and lysosomes), inflamed areas ofthe body (e.g., inflamed joints), tissues, or body compartments. Astable linker may be cleaved by unknown mechanisms. A stable linker mayor may not be cleaved in vivo, but may remain an active agent afterpeptide conjugation.

A peptide and drug conjugated via a linker can be described with theformula Peptide-A-B-C-Drug, wherein the linker is A-B-C. A can be astable amide link such as that formed by reacting an amine on thepeptide with a linker containing a tetrafluorophenyl (TFP) ester or anNHS ester. A can also be a stable carbamate linker such as that formedby reacting an amine on the peptide with an imidazole carbamate activeintermediate formed by reaction of CDI with a hydroxyl on the linker. Acan also be a stable secondary amine linkage such as that formed byreductive alkylation of the amine on the peptide with an aldehyde orketone group on the linker. A can also be a stable thioether linkerformed using a maleimide or bromoacetamide in the linker with a thiol inthe peptide, a triazole linker, a stable oxime linker, or a oxacarbolinelinker. B can be (—CH2—)_(x)— or a short PEG (—CH₂CH₂O—)_(x) (x is 0-20)or other spacers or no spacer. C can be an amide bond formed with anamine or a carboxylic acid on the drug, a thioether formed between amaleimide on the linker and a sulfhydroyl on the drug, a secondary ortertiary amine, a carbamate, or other stable bonds. Any linker chemistrydescribed in “Current ADC Linker Chemistry,” Jain et al., Pharm Res,2015 DOI 10.1007/s11095-015-1657-7 can be used.

The resulting peptide conjugates can be administered to a human oranimal subcutaneously, intravenously, orally, or injected directly intothe kidney to treat disease. The peptide may not be specifically cleavedfrom the detectable agent or active agent via a targeted mechanism. Thepeptide can be degraded by mechanisms such as catabolism, releasing adrug that is modified or not modified form its native form(Antibody-Drug Conjugates: Design, Formulation, and PhysicochemicalStability, Singh, Luisi, and Pak. Pharm Res (2015) 32:3541-3571). Thepeptide drug conjugate exerts its pharmacological activity while stillintact, or while partially or fully degraded, metabolized, orcatabolized.

In some embodiments, peptide conjugates can have cleavable linkers. Insome embodiments, a peptide and drug can be conjugated via a linker andcan be described with the formula Peptide-A-B-C-Drug, wherein the linkeris A-B-C. In some embodiments, A can be a stable amide link such as thatformed by reacting an amine on the peptide with a linker containing atetrafluorophenyl (TFP) ester or an NHS ester. In certain embodiments, Acan also be a stable carbamate linker such as that formed by reacting anamine on the peptide with an imidazole carbamate active intermediateformed by reaction of CDI with a hydroxyl on the linker. In otherembodiments, A can also be a stable secondary amine linkage such as thatformed by reductive alkylation of the amine on the peptide with analdehyde or ketone group on the linker. In some embodiments, A can alsobe a stable thioether linker formed using a maleimide or bromoacetamidein the linker with a thiol in the peptide, a triazole linker, a stableoxime linker, or an oxacarboline linker. B can be (—CH2—)_(x)— or ashort PEG (—CH₂CH₂O—)_(x) (x is 0-20) or other spacers or no spacer. Ccan be an ester bond to the hydroxyl or carboxylic acid on the drug, ora carbonate, hydrazone, or acylhydrazone, designed for hydrolyticcleavage. The hydrolytic rate of cleavage can be varied by varying thelocal environment around the bond, including carbon length (—CH2—)_(x),steric hindrance (including adjacent side groups such as methyl, ethyl,cyclic), hydrophilicity or hydrophobicity. In some embodiments, peptideconjugates can have a linear or cyclic ester linkage, which can includeor do not include side chains such as methyl or ethyl groups. A linearester linkage can be more susceptible to cleavage (such as byhydrolysis, an enzyme such as esterase, or other chemical reaction) thana cyclic ester due to steric hindrance or hydrophobicity/hydrophilicityeffects. Likewise, side chains such as methyl or ethyl groups on thelinear ester linkage can optionally make the linkage less susceptible tocleavage than without the side chains. In some embodiments, hydrolysisrate can be affected by local pH, such as lower pH in certaincompartments of the body or of the cell such as endosomes and lysosomesor diseased tissues. In some embodiments, C can also be a pH sensitivegroup such as a hydrazone or oxime linkage. In other embodiments, C canbe a disulfide bond designed to be released by reduction, such as byglutathione. In other embodiments, (or A-B-C) can be a peptidic linkagedesign for cleavable by enzymes. Optionally, a self-immolating groupsuch as pABC can be included to cause release of a free unmodified drugupon cleavage (Antibody-Drug Conjugates: Design, Formulation, andPhysicochemical Stability, Singh, Luisi, and Pak. Pharm Res (2015)32:3541-3571). The linker can be cleaved by enzymes such as esterases,matrix metalloproteinases, cathepsins such as cathepsin B,glucuronidases, a protease, or thrombin. Alternatively, the bonddesigned for cleavage can be at A, rather than C, and C can be a stablebond or a cleavable bond. An alternative design can be to have stablelinkers (such as amide or carbamate) at A and C and have a cleavablelinker in B, such as a disulfide bond. The rate of reduction can bemodulated by local effects such as steric hindrance from methyl or ethylgroups or modulating hydrophobicity/hydrophilicity. In some embodiments,peptide conjugates can have an ester carbonyl linkage, a longhydrocarbon linker, or carbamate linker, each of which can includehydrophilic groups, such as alcohols, acids, or ethers, or can include ahydrocarbon side chain or other moiety that tunes the rate of cleavage.For example, the rate of hydrolysis can be faster with hydrophilicgroups, such as alcohols, acids, or ethers, near an ester carbonyl. Inanother example, hydrophobic groups present as side chains or as alonger hydrocarbon linker can slow the cleavage rate of the ester.Likewise, cleavage of a carbamate group can also be tuned by hindrance,hydrophobicity, and the like. In another example, using a less labilelinking group, such as a carbamate rather than an ester, can slow thecleavage rate of the linker.

Non-limiting examples of linkers include:

wherein each n is independently 0 to about 1,000; 1 to about 1,000; 0 toabout 500; 1 to about 500; 0 to about 250; 1 to about 250; 0 to about200; 1 to about 200; 0 to about 150; 1 to about 150; 0 to about 100; 1to about 100; 0 to about 50; 1 to about 50; 0 to about 40; 1 to about40; 0 to about 30; 1 to about 30; 0 to about 25; 1 to about 25; 0 toabout 20; 1 to about 20; 0 to about 15; 1 to about 15; 0 to about 10; 1to about 10; 0 to about 5; or 1 to about 5. In some embodiments, each nis independently 0, about 1, about 2, about 3, about 4, about 5, about6, about 7, about 8, about 9, about 10, about 11, about 12, about 13,about 14, about 15, about 16, about 17, about 18, about 19, about 20,about 21, about 22, about 23, about 24, about 25, about 26, about 27,about 28, about 29, about 30, about 31, about 32, about 33, about 34,about 35, about 36, about 37, about 38, about 39, about 40, about 41,about 42, about 43, about 44, about 45, about 46, about 47, about 48,about 49, or about 50. In some embodiments, m is 1 to about 1,000; 1 toabout 500; 1 to about 250; 1 to about 200; 1 to about 150; 1 to about100; 1 to about 50; 1 to about 40; 1 to about 30; 1 to about 25; 1 toabout 20; 1 to about 15; 1 to about 10; or 1 to about 5. In someembodiments, m is 0, about 1, about 2, about 3, about 4, about 5, about6, about 7, about 8, about 9, about 10, about 11, about 12, about 13,about 14, about 15, about 16, about 17, about 18, about 19, about 20,about 21, about 22, about 23, about 24, about 25, about 26, about 27,about 28, about 29, about 30, about 31, about 32, about 33, about 34,about 35, about 36, about 37, about 38, about 39, about 40, about 41,about 42, about 43, about 44, about 45, about 46, about 47, about 48,about 49, or about 50.

In some cases a linker can be a succinic linker, and a drug can beattached to a peptide via an ester bond or an amide bond with twomethylene carbons in between. In other cases, a linker can be any linkerwith both a hydroxyl group and a carboxylic acid, such as hydroxyhexanoic acid or lactic acid.

The linker can be a cleavable or a stable linker. The use of a cleavablelinker permits release of the conjugated moiety (e.g., a therapeuticagent) from the peptide, e.g., after targeting to the kidney. In somecases the linker is enzyme cleavable, e.g., a valine-citrulline linker.In some embodiments, the linker contains a self-immolating portion. Inother embodiments, the linker includes one or more cleavage sites for aspecific protease, such as a cleavage site for matrix metalloproteases(MMPs), thrombin, or cathepsin. Alternatively or in combination, thelinker is cleavable by other mechanisms, such as via pH, reduction,thiol exchange or hydrolysis. The use of a cleavable linker permitsrelease of the conjugated moiety (e.g., a therapeutic agent) from thepeptide, e.g., after targeting to the renal tissue. A hydrolyticallylabile linker, (amongst other cleavable linkers described herein) can beadvantageous in terms of releasing active agents from the peptide. Forexample, an active agent in a conjugate form with the peptide may not beactive, but upon release from the conjugate after targeting to thekidney, the active agent is active. Alternatively, a stable linker canstill permit release of an active cleavage product after catabolism in acell. In some embodiments, a peptide can be conjugated to an activeagent by common techniques known in the art, such those described inBioconjugate Techniques by Greg T. Hermanson (Elsevier Inc., 3^(rd)Edition, 2013)).

The rate of hydrolysis of the linker can be tuned. For example, the rateof hydrolysis of linkers with unhindered esters is faster compared tothe hydrolysis of linkers with bulky groups next an ester carbonyl. Asadditional examples, the rate of disulfide cleavage or exchange withunhindered disulfides is faster compared to the rate of disulfidecleavage or exchange of linkers with bulky groups near disulfide bonds.Protease sites can also affect cleavage rates. A bulky group can be amethyl group, an ethyl group, a phenyl group, a ring, or an isopropylgroup, or any group that provides steric bulk. In some cases, the stericbulk can be provided by the drug itself, such as by ketorolac whenconjugated via its carboxylic acid. The rate of hydrolysis of the linkercan be tuned according to the residency time of the conjugate in thekidney. For example, when a peptide is cleared from the kidneyrelatively quickly, the linker can be tuned to rapidly hydrolyze. Incontrast, for example, when a peptide has a longer residence time in thekidney, a slower hydrolysis rate can allow for extended delivery of anactive agent. This can be important when the peptide is used to delivera drug to the kidney. “Programmed hydrolysis in designing paclitaxelprodrug for nanocarrier assembly” Sci Rep 2015, 5, 12023 Fu et al.,provides an example of modified hydrolysis rates.

Peptide Stability

A peptide of the present disclosure can be stable in various biologicalconditions as well as during manufacturing, handling, storage, and otherconditions in either a liquid or a dried state. Additionally, a peptideof the present disclosure can be resistant to enzymatic cleavage neededfor peptide processing by the immune system. For example, any peptide ofSEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO:213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 569 can exhibit resistance toreducing agents, proteases, oxidative conditions, or acidic conditions.

In some cases, biologic molecules (such as peptides and proteins) canprovide therapeutic functions, but such therapeutic functions aredecreased or impeded by instability caused by the in vivo environment.(Moroz et al. Adv Drug Deliv Rev 101:108-21 (2016), Mitragotri et al.Nat Rev Drug Discov 13(9):655-72 (2014), Bruno et al. Ther Deliv(11):1443-67 (2013), Sinha et al. Crit Rev Ther Drug Carrier Syst.24(1):63-92 (2007), Hamman et al. BioDrugs 19(3):165-77 (2005)).

Peptide degradation can be a result of a number of processes involvinghydrolytic pathways, peptide oxidation such as oxidation of methionine(Met) residues, deamidation of asparagine (Asn) and glutamine (Gln)residues, and isomerization and hydrolysis of an adjacent asparagine(Asp) residue. (Manning et al., Pharmaceutical Research, Vol. 27 No. 4(2010)). The amino acid immediately following the Asn or Gln residue canalso affect the rate of deamidation, whereas: Asn-Gly, Asn-Ser, Asn-His,and Gln-Gly can be more likely to undergo deamidation. Additionally, thepeptide bond adjacent to amino acids such as Asp can undergo hydrolysiswith amino acid pairings such as Asp-Gly, Asp-Ser, Asp-Tyr, and Asp-Pro,which can be more likely to undergo hydrolysis. Oxidation of amino acidresidues such as Met can form a sulfoxide species. The specificdegradation reactions rates can vary for any given peptide or proteinsequence.

Furthermore, the microenvironment within the molecular structure of thepeptide, solvent accessibility, and conformational stability of eachresidue can impact the likelihood of peptide degradation. Therefore, bymodifying a peptide sequence to reduce occurrence of such degradationevents, a the modified peptide or peptide-conjugate can have increasedbeneficial properties over unmodified peptides or peptide-drugconjugates, such as improved therapeutic efficacy, an increased safetyprofile, and can be less expensive to manufacture and develop. Keyformulaic considerations that can prevent peptide decay can include theuse of excipients, formulation at a desired pH, and storage underspecific conditions (e.g., temperature, oxygen, light exposure, solid orliquid state, and container excipient materials). To circumventdegradation, peptide residues can be substituted with amino acids thatincrease stability, which can result in more efficacious and durabletherapeutic peptides.

With respect to in vivo stability, the GI tract can contain a region oflow pH (e.g. pH˜1), a reducing environment, or a protease-richenvironment that can degrade peptides and proteins. Proteolytic activityin other areas of the body, such as the mouth, eye, lung, intranasalcavity, skin, vaginal tract, mucous membranes, and serum, can also be anobstacle to the delivery of functionally active peptides andpolypeptides. Additionally, the half-life of peptides in serum can bevery short, in part due to proteases, such that the peptide can bedegraded too quickly to have a lasting therapeutic effect whenadministering a therapeutic and safe dosing regimen. Likewise,proteolytic activity in cellular compartments such as lysosomes andreduction activity in lysosomes and the cytosol can degrade peptides andproteins such that they may be unable to provide a therapeutic functionon intracellular targets. Therefore, peptides that are resistant toreducing agents, proteases, and low pH may be able to provide enhancedtherapeutic effects or enhance the therapeutic efficacy of co-formulatedor conjugated active agents in vivo.

Additionally, oral delivery of drugs can be desirable in order to targetcertain areas of the body (e.g., disease in the GI tract such as coloncancer, irritable bowel disorder, infections, metabolic disorders, andconstipation) despite the obstacles to the delivery of functionallyactive peptides and polypeptides presented by this method ofadministration. For example, oral delivery of drugs can increasecompliance by providing a dosage form that is more convenient forpatients to take as compared to parenteral delivery. Oral delivery canbe useful in treatment regimens that have a large therapeutic window.Therefore, peptides that are resistant to reducing agents, proteases,and low pH can allow for oral delivery of peptides without nullifyingtheir therapeutic function.

Peptide Resistance to Reducing Agents. In some embodiments, a peptide ofthe present disclosure can be reduction resistant. Peptides of thisdisclosure can contain one or more cysteines, which can participate indisulfide bridges that can be integral to preserving the folded state ofthe peptide. Exposure of peptides to biological environments withreducing agents can result in unfolding of the peptide and loss offunctionality and bioactivity. For example, glutathione (GSH) is areducing agent that can be present in many areas of the body and incells, and can reduce disulfide bonds. As another example, a peptide canbecome reduced upon cellular internalization during trafficking of apeptide across the gastrointestinal epithelium after oraladministration. A peptide can become reduced upon exposure to variousparts of the GI tract. The GI tract can be a reducing environment, whichcan inhibit the ability of therapeutic molecules with disulfide bonds tohave optimal therapeutic efficacy, due to reduction of the disulfidebonds. A peptide can also be reduced upon entry into a cell, such asafter internalization by endosomes or lysosomes or into the cytosol, orother cellular compartments. Reduction of the disulfide bonds andunfolding of the peptide can lead to loss of functionality or affect keypharmacokinetic parameters such as bioavailability, peak plasmaconcentration, bioactivity, and half-life. Reduction of the disulfidebonds can also lead to increased susceptibility of the peptide tosubsequent degradation by proteases, resulting in rapid loss of intactpeptide after administration. In some embodiments, a peptide that isresistant to reduction can remain intact and can impart a functionalactivity for a longer period of time in various compartments of the bodyand in cells, as compared to a peptide that is more readily reduced.

In certain embodiments, the peptides of this disclosure can be analyzedfor the characteristic of resistance to reducing agents to identifystable peptides. In some embodiments, the peptides of this disclosurecan remain intact after being exposed to different molarities ofreducing agents such as 0.00001M-0.0001M, 0.0001M-0.001M, 0.001M-0.01M,0.01 M-0.05 M, 0.05 M-0.1 M, for greater 15 minutes or more. In someembodiments, the reducing agent used to determine peptide stability canbe dithiothreitol (DTT), Tris(2-carboxyethyl)phosphine HCl (TCEP),2-Mercaptoethanol, (reduced) glutathione (GSH), or any combinationthereof. In some embodiments, at least 5%-10%, at least 10%-20%, atleast 20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, atleast 60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%-100%of the peptide remains intact after exposure to a reducing agent.

Peptide Resistance to Proteases. In some embodiments, a peptide of thepresent disclosure can be resistant to protease degradation. Thestability of peptides of this disclosure can be determined by resistanceto degradation by proteases. Proteases, also referred to as peptidasesor proteinases, can be enzymes that can degrade peptides and proteins bybreaking bonds between adjacent amino acids. Families of proteases withspecificity for targeting specific amino acids can include serineproteases, cysteine proteases, threonine proteases, aspartic proteases,glutamic proteases, esterases, serum proteases, and asparagineproteases. Additionally, metalloproteases, matrix metalloproteases,elastase, carboxypeptidases, Cytochrome P450 enzymes, and cathepsins canalso digest peptides and proteins. Proteases can be present at highconcentration in blood, in mucous membranes, lungs, skin, the GI tract,the mouth, nose, eye, and in compartments of the cell. Misregulation ofproteases can also be present in various diseases such as rheumatoidarthritis and other immune disorders. Degradation by proteases canreduce bioavailability, biodistribution, half-life, and bioactivity oftherapeutic molecules such that they are unable to perform theirtherapeutic function. In some embodiments, peptides that are resistantto proteases can better provide therapeutic activity at reasonablytolerated concentrations in vivo.

In some embodiments, peptides of this disclosure can resist degradationby any class of protease. In certain embodiments, peptides of thisdisclosure resist degradation by pepsin (which can be found in thestomach), trypsin (which can be found in the duodenum), serum proteases,or any combination thereof. In certain embodiments, peptides of thisdisclosure can resist degradation by lung proteases (e.g., serine,cysteinyl, and aspartyl proteases, metalloproteases, neutrophilelastase, alpha-1 antitrypsin, secretory leucoprotease inhibitor,elafin), or any combination thereof. In some embodiments, the proteasesused to determine peptide stability can be pepsin, trypsin,chymotrypsin, or any combination thereof. In some embodiments, at least5%-10%, at least 10%-20%, at least 20%-30%, at least 30%-40%, at least40%-50%, at least 50%-60%, at least 60%-70%, at least 70%-80%, at least80%-90%, or at least 90%-100% of the peptide remains intact afterexposure to a protease. Peptides of SEQ ID NO: 231, SEQ ID NO: 45, andSEQ ID NO: 132 can have particular structural qualities, which make themmore resistant to protease degradation. For example, peptide of SEQ IDNO: 45 and SEQ ID NO: 133 exhibit the “hitchin” topology as describedpreviously, which can be associated with resistance to protease andchemical degradation.

Peptide Stability in Acidic Conditions. Peptides of this disclosure canbe administered in biological environments that are acidic. For example,after oral administration, peptides can experience acidic environmentalconditions in the gastric fluids of the stomach and gastrointestinal(GI) tract. The pH of the stomach can range from ˜1-4 and the pH of theGI tract ranges from acidic to normal physiological pH descending fromthe upper GI tract to the colon. In addition, the vagina, lateendosomes, and lysosomes can also have acidic pH values, such as lessthan pH 7. The pH of various compartments of the kidney can also vary.These acidic conditions can lead to denaturation of peptides andproteins into unfolded states. Unfolding of peptides and proteins canlead to increased susceptibility to subsequent digestion by otherenzymes as well as loss of biological activity of the peptide.

In certain embodiments, the peptides of this disclosure can resistdenaturation and degradation in acidic conditions and in buffers, whichsimulate acidic conditions. In certain embodiments, peptides of thisdisclosure can resist denaturation or degradation in buffer with a pHless than 1, a pH less than 2, a pH less than 3, a pH less than 4, a pHless than 5, a pH less than 6, a pH less than 7, or a pH less than 8. Insome embodiments, peptides of this disclosure remain intact at a pH of1-3. In certain embodiments, at least 5%-10%, at least 10%-20%, at least20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, at least60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%-100% of thepeptide remains intact after exposure to a buffer with a pH less than 1,a pH less than 2, a pH less than 3, a pH less than 4, a pH less than 5,a pH less than 6, a pH less than 7, or a pH less than 8. In otherembodiments, at least 5%-10%, at least 10%-20%, at least 20%-30%, atleast 30%-40%, at least 40%-50%, at least 50%-60%, at least 60%-70%, atleast 70%-80%, at least 80%-90%, or at least 90%-400% of the peptideremains intact after exposure to a buffer with a pH of 1-3. In otherembodiments, the peptides of this disclosure can be resistant todenaturation or degradation in simulated gastric fluid (pH 1-2). In someembodiments, at least 5-10%, at least 10%-20%, at least 20%-30%, atleast 30%-40%, at least 40%-50%, at least 50%-60%, at least 60%-70%, atleast 70%-80%, at least 80%-90%, or at least 90-100% of the peptideremains intact after exposure to simulated gastric fluid. In someembodiments, low pH solutions such as simulated gastric fluid or citratebuffers can be used to determine peptide stability.

Peptide Stability at High Temperatures. In some embodiments, thepeptides of the present disclosure are resistant to an elevatedtemperature. Peptides of this disclosure can be administered inbiological environments with high temperatures. For example, after oraladministration, peptides can experience high temperatures in the body.Body temperature can range from 36° C. to 40° C. High temperatures canlead to denaturation of peptides and proteins into unfolded states.Unfolding of peptides and proteins can lead to increased susceptibilityto subsequent digestion by other enzymes as well as loss of biologicalactivity of the peptide. In some embodiments, a peptide of thisdisclosure can remain intact at temperatures from 25° C. to 100° C. Hightemperatures can lead to faster degradation of peptides. Stability at ahigher temperature can allow for storage of the peptide in tropicalenvironments or areas where access to refrigeration is limited. Incertain embodiments, 5%-100% of the peptide can remain intact afterexposure to 25° C. for 6 months to 5 years. 5%-100% of a peptide canremain intact after exposure to 70° C. for 15 minutes to 1 hour. 5%-100%of a peptide can remain intact after exposure to 100° C. for 15 minutesto 1 hour. In other embodiments, at least 5%-10%, at least 10%-20%, atleast 20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, atleast 60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%-100%of the peptide remains intact after exposure to 25° C. for 6 months to 5years. In other embodiments, at least 5%-10%, at least 10%-20%, at least20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, at least60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%-100% of thepeptide remains intact after exposure to 70° C. for 15 minutes to 1hour. In other embodiments, at least 5%-10%, at least 10%-20%, at least20%-30%, at least 30%-40%, at least 40%-50%, at least 50%-60%, at least60%-70%, at least 70%-80%, at least 80%-90%, or at least 90%-100% of thepeptide remains intact after exposure to 100° C. for 15 minutes to 1hour.

In some embodiments, the peptide of the peptide active agent conjugatecomprises a sequence that has at least 75%, at least 80%, at least 85%,at least 90%, at least 95%, at least 97%, at least 99% or 100% sequenceidentity with any one of SEQ ID NO: 1-SEQ ID NO: 41 or a fragmentthereof. In some embodiments, the peptide of the peptide active agentconjugate comprises a sequence that has at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, at least 97%, at least 99% or100% sequence identity with any one of SEQ ID NO: 42-SEQ ID NO: 120, SEQID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, or SEQ ID NO: 216-SEQ ID NO:235, or a fragment thereof. In some embodiments, the peptide of thepeptide active agent conjugate comprises a sequence that has at least75%, at least 80%, at least 85%, at least 90%, at least 95%, at least97%, at least 99% or 100% sequence identity with any one of SEQ ID NO:236-SEQ ID NO: 276 or a fragment thereof. In some embodiments, thepeptide of the peptide active agent conjugate comprises a sequence thathas at least 75%, at least 80%, at least 85%, at least 90%, at least95%, at least 97%, at least 99% or 100% sequence identity with any oneof SEQ ID NO: 277-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ IDNO: 448, or SEQ ID NO: 451-SEQ ID NO: 470, or a fragment thereof. Insome embodiments, the peptide of the peptide active agent conjugatecomprises a sequence that has at least 75%, at least 80%, at least 85%,at least 90%, at least 95%, at least 97%, at least 99% or 100% sequenceidentity with any one of SEQ ID NO: 471-SEQ ID NO: 529 or a fragmentthereof. In some embodiments, the peptide of the peptide active agentconjugate comprises a sequence of any one of SEQ ID NO: 1-SEQ ID NO: 41or a fragment thereof. In some embodiments, the peptide of the peptideactive agent conjugate comprises a sequence of any one of SEQ ID NO:42-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, or SEQID NO: 216-SEQ ID NO: 235, or a fragment thereof. In some embodiments,the peptide of the peptide active agent conjugate comprises a sequenceof any one of SEQ ID NO: 236-SEQ ID NO: 276 or a fragment thereof. Insome embodiments, the peptide of the peptide active agent conjugatecomprises a sequence of any one of SEQ ID NO: 277-SEQ ID NO: 355, SEQ IDNO: 362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO:470, or a fragment thereof. In some embodiments, the peptide of thepeptide active agent conjugate comprises a sequence of any one of SEQ IDNO: 471-SEQ ID NO: 529 or a fragment thereof. In some embodiments, thepeptide of the peptide active agent conjugate comprises a sequence ofany one of SEQ ID NO: 530-SEQ ID NO: 549 or SEQ ID NO: 570, or afragment thereof. In some embodiments, the peptide of the peptide activeagent conjugate comprises a sequence of any one of SEQ ID NO: 550-SEQ IDNO: 569 or a fragment thereof. In some embodiments, the peptidecomprises a sequence that has at least 70%, at least 75%, at least 80%,at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or100% sequence identity with any one of SEQ ID NO: 1-SEQ ID NO: 41 or afragment thereof. In some embodiments, the peptide comprises a sequencethat has at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, at least 97%, at least 99%, or 100% sequenceidentity with any one of SEQ ID NO: 42-SEQ ID NO: 120, SEQ ID NO:127-SEQ ID NO: 206, SEQ ID NO: 213, or SEQ ID NO: 216-SEQ ID NO: 235, ora fragment thereof. In some embodiments, the peptide comprises asequence that has at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100%sequence identity with any one of SEQ ID NO: 236-SEQ ID NO: 276 or afragment thereof. In some embodiments, the peptide comprises a sequencethat has at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, at least 97%, at least 99%, or 100% sequenceidentity with any one of SEQ ID NO: 277-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 470, ora fragment thereof. In some embodiments, the peptide comprises asequence that has at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100%sequence identity with any one of SEQ ID NO: 471-SEQ ID NO: 529 or afragment thereof. In some embodiments, the peptide comprises a sequenceof any one of SEQ ID NO: 1-SEQ ID NO: 41 or a fragment thereof. In someembodiments, the peptide comprises a sequence of any one of SEQ ID NO:42-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, or SEQID NO: 216-SEQ ID NO: 235, or a fragment thereof. In some embodiments,the peptide comprises a sequence of any one of SEQ ID NO: 236-SEQ ID NO:276 or a fragment thereof. In some embodiments, the peptide comprises asequence of any one of SEQ ID NO: 277-SEQ ID NO: 355, SEQ ID NO: 362-SEQID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 470, or afragment thereof. In some embodiments, the peptide comprises a sequenceof any one of SEQ ID NO: 471-SEQ ID NO: 529 or a fragment thereof. Insome embodiments, the peptide comprises a sequence of any one of SEQ IDNO: 530-SEQ ID NO: 549 or SEQ ID NO: 570, or a fragment thereof. In someembodiments, the peptide comprises a sequence of any one of SEQ ID NO:550-SEQ ID NO: 569 or a fragment thereof. In some embodiments, thepeptide active agent conjugate or the peptide comprises a peptide withat least 30%, at least 40%, at least 50%, at least 60%, at least 70%, atleast 80%, at least 90%, at least 95%, at least, 97%, at least 98%, orat least 99% identical to any one of SEQ ID NO: 1-SEQ ID NO: 120, SEQ IDNO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355,SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ IDNO: 529.

Pharmacokinetics of Peptides

The pharmacokinetics of any of the peptides of this disclosure can bedetermined after administration of the peptide via different routes ofadministration. For example, the pharmacokinetic parameters of a peptideof this disclosure can be quantified after intravenous, subcutaneous,intramuscular, rectal, aerosol, parenteral, ophthalmic, pulmonary,transdermal, vaginal, optic, nasal, oral, sublingual, inhalation,dermal, intrathecal, intranasal, intra-articular, peritoneal, buccal,synovial, or topical administration. Peptides of the present disclosurecan be analyzed by using tracking agents such as radiolabels orfluorophores. For example, a radiolabeled peptides of this disclosurecan be administered via various routes of administration. Peptideconcentration or dose recovery in various biological samples such asplasma, urine, feces, any organ, skin, muscle, and other tissues can bedetermined using a range of methods including HPLC, fluorescencedetection techniques (TECAN quantification, flow cytometry, iVIS), orliquid scintillation counting.

The methods and compositions described herein can relate topharmacokinetics of peptide administration via any route to a subject.Pharmacokinetics can be described using methods and models, for example,compartmental models or noncompartmental methods. Compartmental modelsinclude but are not limited to monocompartmental model, the twocompartmental model, the multicompartmental model or the like. Modelscan be divided into different compartments and can be described by thecorresponding scheme. For example, one scheme is the absorption,distribution, metabolism and excretion (ADME) scheme. For anotherexample, another scheme is the liberation, absorption, distribution,metabolism and excretion (LADME) scheme. In some aspects, metabolism andexcretion can be grouped into one compartment referred to as theelimination compartment. For example, liberation can include liberationof the active portion of the composition from the delivery system,absorption includes absorption of the active portion of the compositionby the subject, distribution includes distribution of the compositionthrough the blood plasma and to different tissues, metabolism, whichincludes metabolism or inactivation of the composition and finallyexcretion, which includes excretion or elimination of the composition orthe products of metabolism of the composition. Compositions administeredintravenously to a subject can be subject to multiphasic pharmacokineticprofiles, which can include but are not limited to aspects of tissuedistribution and metabolism/excretion. As such, the decrease in plasmaor serum concentration of the composition is often biphasic, including,for example an alpha phase and a beta phase, occasionally a gamma, deltaor other phase is observed

Pharmacokinetics includes determining at least one parameter associatedwith administration of a peptide to a subject. In some aspects,parameters include at least the dose (D), dosing interval (τ), areaunder curve (AUC), maximum concentration (C_(max)), minimumconcentration reached before a subsequent dose is administered(C_(min)), minimum time (T_(min)), maximum time to reach Cmax (T_(max)),volume of distribution (V_(d)), steady-state volume of distribution(V_(ss)), back-extrapolated concentration at time 0 (C₀), steady stateconcentration (C_(ss)), elimination rate constant (k_(e)), infusion rate(k_(in)), clearance (CL), bioavailability (f), fluctuation (% PTF) andelimination half-life (t_(1/2)).

In certain embodiments, the peptides of any of SEQ ID NO: 1-SEQ ID NO:569 exhibit optimal pharmacokinetic parameters after oraladministration. In other embodiments, the peptides of any of SEQ ID NO:1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ IDNO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448,or SEQ ID NO: 451-SEQ ID NO: 569 exhibit optimal pharmacokineticparameters after any route of administration, such as oraladministration, inhalation, intranasal administration, topicaladministration, parenteral administration, intravenous administration,subcutaneous administration, intra-articular administration,intramuscular administration, intraperitoneal administration,transdermal administration, dermal administration, or any combinationthereof.

In some embodiments any peptide of SEQ ID NO: 1-SEQ ID NO: 120, SEQ IDNO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355,SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ IDNO: 569 exhibits an average T_(max) of 0.5-12 hours, or 1-48 hours atwhich the C_(max) is reached, an average bioavailability in serum of0.1%-10% in the subject after administering the peptide to the subjectby an oral route, an average bioavailability in serum of less than 0.1%after oral administration to a subject for delivery to the GI tract, anaverage bioavailability in serum of 10-100% after parenteraladministration, an average t½ of 0.1 hours-168 hours, or 0.25 hours-48hours in a subject after administering the peptide to the subject, anaverage clearance (CL) of 0.5-100 L/hour or 0.5-50 L/hour of the peptideafter administering the peptide to a subject, an average volume ofdistribution (V_(d)) of 200-20,000 mL in the subject after systemicallyadministering the peptide to the subject, or optionally no systemicuptake, any combination thereof.

Methods of Manufacture

Various expression vector/host systems can be utilized for theproduction of the recombinant expression of peptides described herein.Non-limiting examples of such systems include microorganisms such asbacteria transformed with recombinant bacteriophage DNA, plasmid DNA orcosmid DNA expression vectors containing a nucleic acid sequenceencoding peptides or peptide fusion proteins/chimeric proteins describedherein, yeast transformed with recombinant yeast expression vectorscontaining the aforementioned nucleic acid sequence, insect cell systemsinfected with recombinant virus expression vectors (e.g., baculovirus)containing the aforementioned nucleic acid sequence, plant cell systemsinfected with recombinant virus expression vectors (e.g., cauliflowermosaic virus (CaMV), tobacco mosaic virus (TMV) or transformed withrecombinant plasmid expression vectors (e.g., Ti plasmid) containing theaforementioned nucleic acid sequence, or animal cell systems infectedwith recombinant virus expression vectors (e.g., adenovirus, vacciniavirus) including cell lines engineered to contain multiple copies of theaforementioned nucleic acid sequence, either stably amplified (e.g.,CHO/dhfr, CHO/glutamine synthetase) or unstably amplified indouble-minute chromosomes (e.g., murine cell lines). Disulfide bondformation and folding of the peptide could occur during expression orafter expression or both.

A host cell can be adapted to express one or more peptides describedherein. The host cells can be prokaryotic, eukaryotic, or insect cells.In some cases, host cells are capable of modulating the expression ofthe inserted sequences, or modifying and processing the gene or proteinproduct in the specific fashion desired. For example, expression fromcertain promoters can be elevated in the presence of certain inducers(e.g., zinc and cadmium ions for metallothionine promoters). In somecases, modifications (e.g., phosphorylation) and processing (e.g.,cleavage) of peptide products can be important for the function of thepeptide. Host cells can have characteristic and specific mechanisms forthe post-translational processing and modification of a peptide. In somecases, the host cells used to express the peptides secretes minimalamounts of proteolytic enzymes.

In the case of cell- or viral-based samples, organisms can be treatedprior to purification to preserve and/or release a target polypeptide.In some embodiments, the cells are fixed using a fixing agent. In someembodiments, the cells are lysed. The cellular material can be treatedin a manner that does not disrupt a significant proportion of cells, butwhich removes proteins from the surface of the cellular material, and/orfrom the interstices between cells. For example, cellular material canbe soaked in a liquid buffer or, in the case of plant material, can besubjected to a vacuum, in order to remove proteins located in theintercellular spaces and/or in the plant cell wall. If the cellularmaterial is a microorganism, proteins can be extracted from themicroorganism culture medium. Alternatively, the peptides can be packedin inclusion bodies. The inclusion bodies can further be separated fromthe cellular components in the medium. In some embodiments, the cellsare not disrupted. A cellular or viral peptide that is presented by acell or virus can be used for the attachment and/or purification ofintact cells or viral particles. In addition to recombinant systems,Peptides can also be synthesized in a cell-free system using a varietyof known techniques employed in protein and peptide synthesis.

In some cases, a host cell produces a peptide that has an attachmentpoint for a drug. An attachment point could comprise a lysine residue,an N-terminus, a cysteine residue, a cysteine disulfide bond, or anon-natural amino acid. The peptide could also be producedsynthetically, such as by solid-phase peptide synthesis, orsolution-phase peptide synthesis. Peptide synthesis can also beperformed by fluorenylmethyloxycarbonyl (Fmoc) chemistry or bybutyloxycarbonyl (Boc) chemistry. The peptide could be folded (formationof disulfide bonds) during synthesis or after synthesis or both. Peptidefragments could be produced enzymatically or synthetically orrecombinantly and then joined together synthetically, recombinantly, orvia an enzyme.

FIG. 4 illustrates a schematic of a method of manufacturing a constructthat expresses a peptide of the disclosure, such as the constructsillustrated in FIG. 3 and as described throughout the disclosure and inSEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO:213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 569 provided herein.

In other aspects, the peptides of the present disclosure can be preparedby conventional solid phase chemical synthesis techniques, for exampleaccording to the Fmoc solid phase peptide synthesis method (“Fmoc solidphase peptide synthesis, a practical approach,” edited by W. C. Chan andP. D. White, Oxford University Press, 2000), by Boc solid phase peptidesynthesis, or by conventional solution phase peptide synthesis.Refolding and disulfide bond formation can be executed by methods knownin the art, such as incubation of the peptide at a mildly basic pH inthe presence of a redox pair such as reduced and oxidized cysteine,either after cleavage and protecting group removal and purification, orwhile still on the resin. Peptide fragments can also be madesynthetically or recombinantly and then joined together. [. Thedisulfide bonds can be formed after cleavage from the resin, such as byair oxidation or a buffer system with a set pH range such as from 7-10and can contain a redox system such as glutathione/oxidized glutathioneor cysteine/cystine. The disulfide bonds can also be formed by selectiveprotection and deprotection of specific cysteine residues followed byoxidation or on the resin. The peptide can be purified, such as byreversed-phase chromatography at any one or more steps during theproduction process. The peptide can be isolated by lyophilization andcan be in various salt forms, such as TFA salt or ammonium and acetatesalt.

Pharmaceutical Compositions of Peptides and Peptide-Conjugates

A pharmaceutical composition of the disclosure can be a combination ofany peptide or peptide-conjugate described herein, or a salt thereof,with other chemical components, such as carriers, stabilizers, diluents,dispersing agents, suspending agents, thickening agents, antioxidants,solubilizers, buffers, osmolytes, salts, surfactants, amino acids,encapsulating agents, bulking agents, cryoprotectants, and/orexcipients. The pharmaceutical composition facilitates administration ofa peptide or peptide-conjugate described herein to an organism.Pharmaceutical compositions can be administered intherapeutically-effective amounts as pharmaceutical compositions byvarious forms and routes including, for example, intravenous,subcutaneous, intramuscular, rectal, aerosol, parenteral, ophthalmic,pulmonary, transdermal, vaginal, optic, nasal, oral, sublingual,inhalation, dermal, intrathecal, intranasal, intra-articular, topicaladministration, or combination thereof. A pharmaceutical composition canbe administered in a local or systemic manner, for example, viainjection of the peptide described herein directly into an organ,optionally in a depot.

Parenteral injections can be formulated for bolus injection orcontinuous infusion. The pharmaceutical compositions can be in a formsuitable for parenteral injection as a sterile suspension, solution oremulsion in oily or aqueous vehicles, and can contain formulatory agentssuch as suspending, stabilizing and/or dispersing agents. Pharmaceuticalformulations for parenteral administration include aqueous solutions ofa peptide described herein in water soluble form. Suspensions ofpeptides described herein can be prepared as oily injection suspensions.Suitable lipophilic solvents or vehicles include fatty oils such assesame oil, or synthetic fatty acid esters, such as ethyl oleate ortriglycerides, or liposomes. Aqueous injection suspensions can containsubstances which increase the viscosity of the suspension, such assodium carboxymethyl cellulose, sorbitol, or dextran. The suspension canalso contain suitable stabilizers or agents which increase thesolubility and/or reduces the aggregation of such peptides describedherein to allow for the preparation of highly concentrated solutions.Alternatively, the peptides described herein can be lyophilized or inpowder form for re-constitution with a suitable vehicle, e.g., sterilepyrogen-free water, before use. In some embodiments, a purified peptideis administered intravenously.

A peptide or peptide-conjugate of the disclosure can be applied directlyto an organ, or an organ tissue or cells, such as brain or brain tissueor cancer cells, during a surgical procedure. The recombinant peptidesdescribed herein can be administered topically and can be formulatedinto a variety of topically administrable compositions, such assolutions, suspensions, lotions, gels, pastes, medicated sticks, balms,creams, and ointments. Such pharmaceutical compositions can containsolubilizers, stabilizers, tonicity enhancing agents, buffers andpreservatives.

In practicing the methods of treatment or use provided herein,therapeutically-effective amounts of the peptide described hereindescribed herein can be administered in pharmaceutical compositions to asubject suffering from a condition that affects the immune system. Insome embodiments, the subject is a mammal such as a human. Atherapeutically-effective amount can vary widely depending on theseverity of the disease, the age and relative health of the subject, thepotency of the compounds used, and other factors.

Pharmaceutical compositions can be formulated using one or morephysiologically-acceptable carriers comprising excipients andauxiliaries, which facilitate processing of the active compounds intopreparations that can be used pharmaceutically. Formulation can bemodified depending upon the route of administration chosen.Pharmaceutical compositions comprising a peptide described herein can bemanufactured, for example, by expressing the peptide in a recombinantsystem, purifying the peptide, lyophilizing the peptide, mixing,dissolving, granulating, dragee-making, levigating, emulsifying,encapsulating, entrapping, or compression processes. The pharmaceuticalcompositions can include at least one pharmaceutically acceptablecarrier, diluent, or excipient and compounds described herein asfree-base or pharmaceutically-acceptable salt form.

Methods for the preparation of peptides described herein comprising thecompounds described herein include formulating the peptide orpeptide-conjugates described herein, or a salt thereof, with one or moreinert, pharmaceutically-acceptable excipients or carriers to form asolid, semi-solid, or liquid composition. Solid compositions include,for example, powders, tablets, dispersible granules, capsules, cachets,and suppositories. These compositions can also contain minor amounts ofnontoxic, auxiliary substances, such as wetting or emulsifying agents,pH buffering agents, and other pharmaceutically-acceptable additives.

Non-limiting examples of pharmaceutically-acceptable excipients can befound, for example, in Remington: The Science and Practice of Pharmacy,Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, JohnE., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton,Pa. 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical DosageForms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams &Wilkins 1999), each of which is incorporated by reference in itsentirety.

Administration of Pharmaceutical Compositions

A pharmaceutical composition of the disclosure can be a combination ofany peptide described herein with other chemical components, such ascarriers, stabilizers, diluents, dispersing agents, suspending agents,thickening agents, and/or excipients. The pharmaceutical compositionfacilitates administration of a peptide described herein to an organism.Pharmaceutical compositions can be administered intherapeutically-effective amounts as pharmaceutical compositions byvarious forms and routes including, for example, intravenous,subcutaneous, intramuscular, rectal, aerosol, parenteral, ophthalmic,pulmonary, transdermal, vaginal, optic, nasal, oral, inhalation, dermal,intra-articular, intrathecal, intranasal, and topical administration. Apharmaceutical composition can be administered in a local or systemicmanner, for example, via injection of the peptide described hereindirectly into an organ, optionally in a depot.

Parenteral injections can be formulated for bolus injection orcontinuous infusion. The pharmaceutical compositions can be in a formsuitable for parenteral injection as a sterile suspension, solution oremulsion in oily or aqueous vehicles, and can contain formulatory agentssuch as suspending, stabilizing and/or dispersing agents. Pharmaceuticalformulations for parenteral administration include aqueous solutions ofa peptide described herein in water-soluble form. Suspensions ofpeptides described herein can be prepared as oily injection suspensions.Suitable lipophilic solvents or vehicles include fatty oils such assesame oil, or synthetic fatty acid esters, such as ethyl oleate ortriglycerides, or liposomes. Aqueous injection suspensions can containsubstances which increase the viscosity of the suspension, such assodium carboxymethyl cellulose, sorbitol, or dextran. The suspension canalso contain suitable stabilizers or agents which increase thesolubility and/or reduce the aggregation of such peptides describedherein to allow for the preparation of highly concentrated solutions.Alternatively, the peptides described herein can be lyophilized or inpowder form for re-constitution with a suitable vehicle, e.g., sterilepyrogen-free water, before use. In some embodiments, a purified peptideis administered intravenously. A peptide described herein can beadministered to a subject, home, target, migrates to, is retained by,and/or binds to, or be directed to an organ, e.g., the kidney.

A peptide of the disclosure can be applied directly to an organ, or anorgan tissue or cells, such as the kidney, kidney tissue, or cells,during a surgical procedure, such as kidney transplantation. Therecombinant peptides described herein can be administered topically andcan be formulated into a variety of topically administrablecompositions, such as solutions, suspensions, lotions, gels, pastes,medicated sticks, balms, creams, and ointments. Such pharmaceuticalcompositions can contain solubilizers, stabilizers, tonicity enhancingagents, buffers and preservatives.

In practicing the methods of treatment or use provided herein,therapeutically-effective amounts of the peptide described hereindescribed herein are administered in pharmaceutical compositions to asubject suffering from a condition. In some instances the pharmaceuticalcomposition will affect the physiology of the animal, such as the immunesystem, inflammatory response, or other physiologic affect. In someembodiments, the subject is a mammal such as a human. Atherapeutically-effective amount can vary widely depending on theseverity of the disease, the age and relative health of the subject, thepotency of the compounds used, and other factors.

Pharmaceutical compositions can be formulated using one or morephysiologically-acceptable carriers comprising excipients andauxiliaries, which facilitate processing of the active compounds intopreparations that can be used pharmaceutically. Formulation can bemodified depending upon the route of administration chosen.Pharmaceutical compositions comprising a peptide described herein can bemanufactured, for example, by expressing the peptide in a recombinantsystem, purifying the peptide, lyophilizing the peptide, mixing,dissolving, granulating, dragee-making, levigating, emulsifying,encapsulating, entrapping, or compression processes. The pharmaceuticalcompositions can include at least one pharmaceutically acceptablecarrier, diluent, or excipient and compounds described herein asfree-base or pharmaceutically-acceptable salt form.

Methods for the preparation of peptides described herein comprising thecompounds described herein include formulating the peptide describedherein with one or more inert, pharmaceutically-acceptable excipients orcarriers to form a solid, semi-solid, or liquid composition. Solidcompositions include, for example, powders, tablets, dispersiblegranules, capsules, cachets, and suppositories. These compositions canalso contain minor amounts of nontoxic, auxiliary substances, such aswetting or emulsifying agents, pH buffering agents, and otherpharmaceutically-acceptable additives.

Non-limiting examples of pharmaceutically-acceptable excipients can befound, for example, in Remington: The Science and Practice of Pharmacy,Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, JohnE., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton,Pa. 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical DosageForms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams &Wilkins 1999), each of which is incorporated by reference in itsentirety.

Use of Peptide in Imaging and Surgical Methods

The present disclosure generally relates to peptides that home, target,migrate to, are retained by, accumulate in, and/or bind to, or aredirected to specific regions, tissues, structures, or cells within thebody and methods of using such peptides. These peptides can contact thekidney, which can make them useful for a variety of applications. Inparticular, the peptides can have applications in site-specificmodulation of biomolecules to which the peptides are directed to. Enduses of such peptides can include, for example, imaging, research,therapeutics, theranostics, pharmaceuticals, chemotherapy, chelationtherapy, targeted drug delivery, and radiotherapy. Some uses can includetargeted drug delivery and imaging.

In some embodiments, the present disclosure provides a method fordetecting a cancer, cancerous tissue, or tumor tissue, the methodcomprising the steps of contacting a tissue of interest with a peptideof the present disclosure, wherein the peptide is conjugated to adetectable agent and measuring the level of binding of the peptide,wherein an elevated level of binding, relative to normal tissue, isindicative that the tissue is a cancer, cancerous tissue or tumortissue.

In some embodiments, the disclosure provides a method of imaging anorgan or body region or region, tissue or structure of a subject, themethod comprising administrating to the subject the peptide or apharmaceutical composition disclosed herein and imaging the subject. Insome embodiments such imaging is used to detect a condition associatedwith a function of the kidney. In some cases the condition is aninflammation, a cancer, a degradation, a growth disturbance, genetic, atear or an injury, or another suitable condition. In some cases thecondition can be Systemic Lupus Erythematosus (SLE or “Lupus”), oranother suitable condition. In some case the condition can be associatedwith a cancer or tumor of the kidney. In some embodiments, such as thoseassociated with cancers, the imaging can be associated with surgicalremoval of the diseased region, tissue, structure or cell of a subject.

Furthermore, the present disclosure provides methods for intraoperativeimaging and resection of a diseased or inflamed tissue, cancer,cancerous tissue, or tumor tissue using a peptide of the presentdisclosure conjugated with a detectable agent. In some embodiments, thediseased or inflamed tissue, cancer, cancerous tissue, or tumor tissueis detectable by fluorescence imaging that allows for intraoperativevisualization of the cancer, cancerous tissue, or tumor tissue using apeptide of the present disclosure. In some embodiments, the peptide ofthe present disclosure is conjugated to one or more detectable agents.In a further embodiment, the detectable agent comprises a fluorescentmoiety coupled to the peptide. In another embodiment, the detectableagent comprises a radionuclide. In some embodiments, imaging is achievedduring open surgery. In further embodiments, imaging is accomplishedusing endoscopy or other non-invasive surgical techniques.

Renal Therapy with Peptides and Peptide-Conjugates

As discussed above and herein, the present disclosure provides peptidesthat home, target, migrate to, accumulate in, are directed to, and/orbind to specific regions, tissues, structures, or cells of the kidneyand methods of using such peptides. End uses of such peptides include,for example, imaging, research, therapeutics, diagnostics, theranostics,pharmaceuticals, chemotherapy, chelation therapy, targeted drugdelivery, and radiotherapy.

In one embodiment, the method includes administering an effective amountof a peptide of the present disclosure to a subject in need thereof. Theterm “effective amount,” as used herein, can refer to a sufficientamount of an agent or a compound being administered which will relieveto some extent one or more of the symptoms of the disease or conditionbeing treated. The result can be reduction and/or alleviation of thesigns, symptoms, or causes of a disease, or any other desired alterationof a biological system. Compositions containing such agents or compoundscan be administered for prophylactic, enhancing, and/or therapeutictreatments. An appropriate “effective” amount in any individual case canbe determined using techniques, such as a dose escalation study.

The methods, compositions, and kits of this disclosure can comprise amethod to prevent, treat, arrest, reverse, or ameliorate the symptoms ofa condition. The treatment can comprise treating a subject (e.g., anindividual, a domestic animal, a wild animal or a lab animal afflictedwith a disease or condition) with a peptide of the disclosure. Intreating a disease, the peptide can contact the kidney of a subject. Thesubject can be a human. A subject can be a human; a non-human primatesuch as a chimpanzee, or other ape or monkey species; a farm animal suchas a cattle, horse, sheep, goat, swine; a domestic animal such as arabbit, dog, and cat; a laboratory animal including a rodent, such as arat, mouse and guinea pig, or the like. A subject can be of any age. Asubject can be, for example, an elderly adult, adult, adolescent,pre-adolescent, child, toddler, infant, or fetus in utero.

Treatment can be provided to the subject before clinical onset ofdisease. Treatment can be provided to the subject after clinical onsetof disease. Treatment can be provided to the subject after 1 day, 1week, 6 months, 12 months, or 2 years or more after clinical onset ofthe disease. Treatment may be provided to the subject for more than 1day, 1 week, 1 month, 6 months, 12 months, 2 years or more afterclinical onset of disease. Treatment may be provided to the subject forless than 1 day, 1 week, 1 month, 6 months, 12 months, or 2 years afterclinical onset of the disease. Treatment can also include treating ahuman in a clinical trial. A treatment can comprise administering to asubject a pharmaceutical composition, such as one or more of thepharmaceutical compositions described throughout the disclosure. Atreatment can comprise a once daily dosing. A treatment can comprisedelivering a peptide of the disclosure to a subject, eitherparenterally, intravenously, subcutaneously, intramuscularly, byinhalation, dermally, intra-articular injection, orally, intrathecally,transdermally, intranasally, via a peritoneal route, or directly, e.g.,via topical, intra-articular injection route or injection route ofapplication. A treatment can comprise administering a peptide-activeagent complex to a subject, either parenterally, intravenously,subcutaneously, intramuscularly, by inhalation, dermally,intra-articular injection, orally, intrathecally, transdermally,intranasally, via a peritoneal route, or directly onto, near or into thekidney, e.g., via topical, intra-articular injection, or injection routeof application or during surgery.

In some embodiments, the present disclosure provides a method fortreating a cancer, the method comprising administering to a subject inneed thereof an effective amount of a peptide of the present disclosure.

In some embodiments, the present disclosure provides a method fortreating a cancer, the method comprising administering to a patient inneed thereof an effective amount of a pharmaceutical compositioncomprising a peptide of the present disclosure and a pharmaceuticallyacceptable carrier.

In some embodiments, the present disclosure provides a method forinhibiting invasive activity of cells, the method comprisingadministering an effective amount of a peptide of the present disclosureto a subject.

In some embodiments, the peptides of the present disclosure areconjugated to one or more therapeutic agents. In further embodiments,the therapeutic agent is a chemotherapeutic, anti-cancer drug, oranti-cancer agent selected from, but are not limited to:anti-inflammatories, such as for example a glucocorticoid, acorticosteroid, a protease inhibitor, such as for example collagenaseinhibitor or a matrix metalloprotease inhibitor (i.e., MMP-13inhibitor), an amino sugar, vitamin (e.g., Vitamin D), and antibiotics,antiviral, or antifungal, a statin, an immune modulator, radioisotopes,toxins, enzymes, sensitizing drugs, nucleic acids, including interferingRNAs, antibodies, anti-angiogenic agents, cisplatin, anti-metabolites,mitotic inhibitors, growth factor inhibitors, paclitaxel, temozolomide,topotecan, fluorouracil, vincristine, vinblastine, procarbazine,decarbazine, altretamine, methotrexate, mercaptopurine, thioguanine,fludarabine phosphate, cladribine, pentostatin, cytarabine, azacitidine,etoposide, teniposide, irinotecan, docetaxel, doxorubicin, daunorubicin,dactinomycin, idarubicin, plicamycin, mitomycin, bleomycin, tamoxifen,flutamide, leuprolide, goserelin, aminogluthimide, anastrozole,amsacrine, asparaginase, mitoxantrone, mitotane and amifostine, andtheir equivalents, as well as photo-ablation. Some of these activeagents induce programmed cell death such as apoptosis in target cellsand thereby improve symptoms or ameliorate disease. Apoptosis can beinduced by many active agents, including, for example,chemotherapeutics, anti-inflammatories, corticosteroids, NSAIDS, tumornecrosis factor alpha (TNF-α) modulators, tumor necrosis factor receptor(TNFR) family modulators. In some embodiments, peptides of thisdisclosure can be used to target active agents to pathways of cell deathor cell killing, such as caspases, apoptsis activators and inhibitors,XBP-1, Bcl-2, Bcl-Xl, Bcl-w, and other disclosed herein. In otherembodiments, the therapeutic agent is any nonsteroidal anti-inflammatorydrug (NSAID). The NSAID can be any heterocyclic acetic acid derivativessuch as ketorolac, indomethacin, etodolac, or tolemetin, any propionicacid derivatives such as naproxen, any enolic acid derivatives, anyanthranilic acid derivatives, any selective COX-2 inhibitors such ascelecoxib, any sulfonanilides, any salicylates, aceclofenac, nabumetone,sulindac, diclofenac, or ibuprofen. In other embodiments, thetherapeutic agent is any steroid, such as dexamethasone, budesonide,beclomethasone monopropionate, desciclesonide, triamcinolone, cortisone,prednisone, rednisolone, triamcinolone hexacetonide, ormethylprednisolone. In other embodiments, the therapeutic agent is apain reliever, such as acetaminophen, opioids, local anesthetics,anti-depressants, glutamate receptor antagonists, adenosine, orneuropeptides. In some embodiments, a treatment consists ofadministering a combination of any of the above therapeutic agents and apeptide conjugate, such as a treatment in which both adexamethasone-peptide conjugate and an NSAID are administered to apatient. Peptides of the current disclosure that target the kidney canbe used to treat the diseases conditions as described herein, forexample, any diseases or conditions including tears, injuries (i.e.,sports injuries), genetic factors, degradation, thinning, inflammation,cancer or any other disease or condition of the kidney or to targettherapeutically-active substances to treat these diseases amongstothers. In some cases, the peptide or peptide-active agent can be usedto target cancer in the kidney, by contacting the kidney and then havingantitumor function, targeted toxicity, inhibiting metastases, etc. Aswell, such peptide or peptide-active agent can be used to label, detect,or image such kidney lesions, including tumors and metastases amongstother lesions, which may be removed through various surgical techniquesor by targeting with peptide-active agents that induce programmed celldeath or kill cells.

Venom or toxin derived peptide(s), peptides, modified peptides, labeledpeptides, peptide-active agent conjugates and pharmaceuticalcompositions described herein can be administered for prophylacticand/or therapeutic treatments. In therapeutic applications, thecomposition can be administered to a subject already suffering from adisease or condition, in an amount sufficient to cure or at leastpartially arrest the symptoms of the disease or condition, or to cure,heal, improve, or ameliorate the condition. Such peptides describedherein can also be administered to prevent (either in whole or in part),lessen a likelihood of developing, contracting, or worsening acondition. Amounts effective for this use can vary based on the severityand course of the disease or condition, previous therapy, the subject'shealth status, weight, response to the drugs, and the judgment of thetreating physician. Venom or toxin derived peptide(s), peptides,modified peptides, labeled peptides, peptide-active agent conjugates andpharmaceutical compositions described herein can allow for targetedhoming of the peptide and local delivery of any conjugate. For example,a peptide conjugated to a steroid allows for local delivery of thesteroid, which is significantly more effective and less toxic thantraditional systemic steroids. A peptide conjugated to an NSAID isanother example. In this case, the peptide conjugated to an NSAID allowsfor local delivery of the NSAID, which allows for administration of alower NSAID dose and is subsequently less toxic. By delivering an activeagent to the kidney, pain relief can be more rapid, may be more longlasting, and can be obtained with a lower systemic dose and off-siteundesired effects than with systemic dosing without targeting.

Peptides of the current disclosure can be used to treat or manage painassociated with a kidney injury or disorder, or any other kidneycondition as described herein. The peptides can be used either directlyor as carriers of active drugs, peptides, or molecules. For example,since ion channels can be associated with pain and can be activated indisease states, peptides that interact with ion channels can be useddirectly to reduce pain. In another embodiment, the peptide isconjugated to an active agent with anti-inflammatory activity, in whichthe peptide acts as a carrier for the local delivery of the active agentto reduce pain.

In some embodiments, the peptides described herein provide a method oftreating a kidney condition of a subject, the method comprisingadministering to the subject a therapeutically-effective amount of apeptide comprising the sequence ofGSGVX¹IX²X³RCX⁴GSRDCX⁵DPCRX⁶X⁷X⁸GX⁹RX¹⁰GRCX¹¹NRRCRCX¹²X¹³X¹⁴X¹⁵ (SEQ IDNO: 570) or fragment thereof, wherein X1, X2, X3, X4, X5, X6, X7, X8,X9, X10, X11, X12, X13, X14 and X15 are each individually any amino acidor amino acid analogue or null. In some embodiments, the peptidesdescribed herein provide a method of treating a kidney condition of asubject, the method comprising administering to the subject a peptide ofany one of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206,SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO:441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 569 or fragmentthereof.

Treatment of Kidney Disorders

In some embodiments, peptides of this disclosure that home, target, aredirected to, migrate to, are retained by, accumulate in, or bind tospecific regions, tissues, structures, or cells of the kidneys can beused to treat a kidney disorder. In other embodiments, peptides are usedin peptide conjugates of the present disclosure to deliver an activeagent for treatment of a kidney disorder.

In some embodiments, the peptides and peptide-conjugates of the presentdisclosure are used to treat a condition of the kidney, or a region,tissue, structure, or cell thereof. In certain embodiments, thecondition is associated with kidney or a function of a subject'skidneys. The present disclosure encompasses various acute and chronicrenal diseases, including glomerular, tubule-interstitial, andmicrovascular diseases. Examples of conditions applicable to the presentdisclosure include but are not limited to: hypertensive kidney damage,acute kidney diseases and disorders (AKD), acute kidney injury (AKI) dueto ischemia-reperfusion injury, drug treatment such as chemotherapy,cardiovascular surgery, surgery, medical interventions or treatment,radiocontrast nephropathy, or induced by cisplatin or carboplatin, whichcan be treated prophylactically, established AKI including ischemicrenal injury, endotoxemia-induced AKI, endotoxemia/sepsis syndrome, orestablished nephrotoxic AKI (e.g. rhabdomyolysis, radiocontrastnephropathy, cisplatin/carboplatin AKI, aminoglycoside nephrotoxicity),end stage renal disease, acute and rapidly progressiveglomerulonephritis, acute presentations of nephrotic syndrome, acutepyelonephritis, acute renal failure, chronic glomerulonephritis, chronicheart failure, chronic interstitial nephritis, graft versus host diseaseafter renal transplant, chronic kidney disease (CKD) such as diabeticnephropathy, hypertensive nephrosclerosis, idiopathic chronicglomerulonephritis (e.g. focal glomerular sclerosis, membranousnephropathy, membranoproliferative glomerulonephritis, minimal changedisease transition to chronic disease, anti-GBM disease, rapidlyprogressive cresentic glomerulonephritis, IgA nephropathy), secondarychronic glomerulonephritis (e.g. systemic lupus, polyarteritis nodosa,scleroderma, amyloidosis, endocarditis), hereditary nephropathy (e.g.polycystic kidney disease, Alport's syndrome), interstitial nephritisinduced by drugs (e.g. Chinese herbs, NSAIDs), multiple myeloma orsarcoid, or renal transplantation such as donor kidney prophylaxis(treatment of donor kidney prior to transplantation), treatment posttransplantation to treat delayed graft function, acute rejection, orchronic rejection, chronic liver disease, chronic pyelonephritis,diabetes, diabetic kidney disease, fibrosis, focal sclerosis, focalsegmental glomerulosclerosis, Goodpasture's disease, hypertensivenephrosclerosis, IgG4-related renal disease, interstitial inflammation,lupus nephritis, nephritic syndrome, partial obstruction of the urinarytract, polycystic kidney disease, progressive renal disease, renal cellcarcinoma, clear cell renal cell carcinoma, papillary renal cellcarincoma, chromophobe renal cell carinoma, kidney cancer, transitionalcell carcinoma, nephroblastoma, renal sarcoma, renal adenoma,oncocytoma, angiomyolipoma, renal fibrosis, kidney stones, hypertension,hypotension, disorders of sodium, water, acid-base, potassium, calcium,magnesium, or phosphate balance, infections, urinary tract infections,kidney failure, hematuria, renal cysts, uremia, shock, uretalobstruction, proteinuria, Fanconi's syndrome, Bartter's syndrome,chronic renal insufficiency, renal fibrosis, and vasculitis. Forexample, in certain embodiments, the peptides and peptide-conjugates ofthe present disclosure are used to reduce acute kidney injury in orderto prevent it from progressing to chronic kidney disease.

Alternatively or in combination, in some embodiments, the peptide andpeptide-conjugates of the present disclosure are used to elicit aprotective response such as ischemic preconditioning and/or acquiredcytoresistance in a kidney of the subject. In some embodiments, ischemicpreconditioning and/or acquired cytoresistance is induced byadministering an agent (e.g., a peptide or peptide-conjugate of thepresent disclosure) that upregulates the expression of protective stressproteins, such as antioxidants, anti-inflammatory proteins, or proteaseinhibitors. In certain embodiments, the induced response protects thekidney by preserving kidney function in whole or in part and/or byreducing injury to renal tissues and cells, e.g., relative to thesituation where no protective response is induced. The peptides andpeptide-conjugates of the present disclosure can provide certainbenefits compared to other agents for inducing ischemic preconditioningand/or acquired cytoresistance, such as a well-defined chemicalstructure and avoidance of low pH precipitation.

In some embodiments, the protective response is induced in order toprotect the kidney or tissues or cells thereof from an injury or insultthat is predicted to occur (e.g., associated with a planned event suchas a medical procedure, is likely to occur due to a condition in thesubject) or has already occurred. In certain embodiments, the inducedresponse prevents or reduces the extent of damage to the kidney ortissues or cells thereof caused by the injury or insult. For instance,in certain embodiments, the peptides and peptide-conjugates induceacquired cytoresistance by activating protective pathways and/orupregulating expression of protective stress proteins. Optionally, thepeptides and peptide-conjugates are capable of inducing such protectiveresponses while causing minimal or no injury to the kidney.

In various embodiments, the injury or insult is associated with one ormore of: surgery, radiocontrast imaging, cardiopulmonary bypass, balloonangioplasty, induced cardiac or cerebral ischemic-reperfusion injury,organ transplantation, sepsis, shock, low blood pressure, high bloodpressure, kidney hypoperfusion, chemotherapy, drug administration,nephrotoxic drug administration, blunt force trauma, puncture, poison,or smoking. For instance, in certain embodiments, the injury or insultis associated with a medical procedure that has been or will beperformed on the subject, such as one or more of: surgery, radiocontrastimaging, cardiopulmonary bypass, balloon angioplasty, induced cardiac orcerebral ischemic-reperfusion injury, organ transplantation,chemotherapy, drug administration, or nephrotoxic drug administration.

In some embodiments, the peptide itself exhibits a renal therapeuticeffect. For example, in certain embodiments, the cystine-dense peptideinteracts with a renal ion channel, inhibits a protease, hasantimicrobial activity, has anticancer activity, has anti-inflammatoryactivity, induces ischemic preconditioning or acquired cytoresistance,or produces a protective or therapeutic effect on a kidney of thesubject, or a combination thereof. Optionally, the renal therapeuticeffect exhibited by the peptide is a renal protective effect or renalprophylactic effect (e.g., ischemic preconditioning or acquiredcytoresistance) that protects the kidney or a tissue or cell thereoffrom an upcoming injury or insult. Such effects based upon the peptidein and of itself can be used to enhance the therapeutic effect of activeagents that may be conjugated, linked, or grafted to the peptidesdisclosed herein.

For example, in certain embodiments, a peptide of the present disclosureactivates protective pathways and/or upregulates expression ofprotective stress proteins in the kidney or tissues or cells thereof. Asanother example, in certain embodiments, a peptide of the presentdisclosure accesses and suppresses intracellular injury pathways. In yetanother example, in certain embodiments, a peptide of the presentdisclosure inhibits interstitial inflammation and prevents renalfibrosis. As a further example, in certain embodiments, a peptide of thepresent disclosure is administered prior to or currently with theadministration of a nephrotoxic agent (e.g., aminoglycoside antibioticssuch as gentamicin and minocycline, chemotherapeutics such as cisplatin,immunoglobulins or fragments thereof, mannitol, NSAIDs such as ketorolacor ibuprofen, cyclosporin, cyclophosphamide, radiocontrast dyes) inorder to minimize its damaging effects, e.g., by blockingmegalin-cubulin binding sites so that the nephrotoxic agent passesthrough the kidneys.

Alternatively or in combination, in some embodiments, the peptide isconjugated to a renal therapeutic agent that exhibits a renaltherapeutic effect. In certain embodiments, the renal therapeutic agentis used to treat a condition of the kidney, or a region, tissue,structure, or cell thereof, such as the conditions provided herein.Examples of such renal therapeutic agents include but are not limitedto: dexamethasone, a steroid, an anti-inflammatory agent, an antioxidant(e.g., glutathione, N acetyl cysteine), deferoxamine, feroxamine, iron,tin, a metal, a metal chelate, ethylene diamine tetraacetic acid (EDTA),an EDTA-Fe complex, dimercaptosuccinic acid (DMSA),2,3-dimercapto-1-propanesulfonic acid (DMPS), penicillamine, anantibiotic such as gentamicin, vancomycin, minocin or mitomyclin, aniron chelator, a porphyrin, hemin, vitamin B12, a chemotherapeutic, anNrf2 pathway activator such as bardoxolone,angiotensin-converting-enzyme (ACE) inhibitors such as ramipril,captopril, lisinopril, benazepril, quinapril, fosinopril, trandolapril,moexipril, enalaprilat, enalapril maleate, or perindopril erbumine,glycine polymers, or a combination thereof. Additional examples of atherapeutic agent that can be conjugated to the peptide can includeQPI-1002, QM56, SVT016426 (QM31), 16/86 (third generation ferrostatin),BASP siRNA, CCX140, BIIB023, CXA-10, alkaline phosphatase, Dnmt1inhibitor, THR-184, lithium, formoterol, IL-22, EPO and EPO derivatives,agents that stimulate erthyropoietin such as epoeitn alfa ordarbepoietin alfa, PDGF inhibitors, CRMD-001, Atrasentan, Tolvaptan,RWJ-676070, Abatacept, Sotatercept, an anti-infective agent, ananti-viral agent, an anti-fungal agent, an aminoglycoside, animmunosuppresant such tacrolimus, mycophenolic acid (e.g., mycophenolatemofetil), cyclosporine A, or azathioprine, a diuretic drug such asthiazides, bemetanide, ethacrynic acid, furosemide, torsemide, glucose,mannitol, amiloride, spironolactone, eplerenone, triamterene, potassiumcanrenoate, bendroflumethiazide, hydrochlorothiazide, vasopressin,amphotericin B, acetazolamide, tovaptan, conivaptan, dopamine,dorzolamide, bendrolumethiazide, hydrochlorothiazide, caffeine,theophylline, or theobromine, a statin, a senolytic such as navitoclaxor obatoclax, a corticosteroid such as prednisone, betamethasone,fludrocortisone, deoxycorticosterone, aldosterone, cortisone,hydrocortisone, belcometasone, dexamethasone, mometasone, fluticasone,prednisolone, methylprednisolone, triamcinolone acetonide ortriamcinolone, a glucocorticoid, a liposome, renin, SGLT2 modulator, orangiotensin.

For example, in some embodiments, a peptide of the present disclosure isconjugated to an anti-inflammatory agent such as dexamethasone in orderto treat lupus affecting the kidney, vasculitis, Goodpasture's disease,focal segmental glomerulosclerosis, nephritic syndrome, or other renaldisorders caused by inflammatory processes. As another example, in someembodiments, a peptide of the present disclosure is conjugated tochemotherapeutic for treating renal cell carcinoma. As a furtherexample, in some embodiments, a peptide of the present disclosure isconjugated to a steroid for treating polycystic renal disease.

In certain embodiments, the renal therapeutic agent is a renalprotective agent or renal prophylactic agent capable of eliciting aprotective response in the kidney upon administration to a subject. Asdiscussed above and herein, the protective response can protect thekidney or a tissue or cell thereof from an upcoming injury or insult.For example, the renal protective agent or renal prophylactic agent canactivate protective pathways and/or upregulate expression of protectivestress proteins in the kidney or tissues or cells thereof. Examples ofsuch renal protective agents and renal prophylactic agents include butare not limited to: dexamethasone, a steroid, an anti-inflammatoryagent, a nonsteroidal anti-inflammatory drug (NSAID) such as ketorolacor ibuprofen, deferoxamine, iron, tin, a metal, a metal chelate,ethylene diamine tetraacetic acid (EDTA), an EDTA-Fe complex,dimercaptosuccinic acid (DMSA), 2,3-dimercapto-1-propanesulfonic acid(DMPS), penicillamine, an antibiotic, an aminoglycoside, an ironchelator, a porphyrin, vitamin B12, or a combination thereof. In someembodiments, the renal protective agent or renal prophylactic agentcomprises complexed or chelated iron, (e.g., via heme, deferoxamine,feroxamine, porphyrin, EDTA, etc.). In such embodiments, thepeptide-conjugate can be used to deliver iron to the renal tissue forkidney preconditioning.

For example, in certain embodiments, a peptide of the present disclosureis conjugated to hemin, which signals through the heat shock/hemereactive element pathway in order to upregulate a set of diversecytoprotective proteins. As another example, in certain embodiments, apeptide of the present disclosure is conjugated to an iron chelate oriron complex in order to deliver iron to the kidney to alter geneexpression profiles and induce expression of cytoprotective proteins.

The peptides of the present disclosure enable specific targeting ofrenal therapeutic agents and other agents to the kidneys, which in someembodiments is beneficial for reducing undesirable effect associatedwith systemic delivery and/or delivery to non-target tissues. Forexample, patients with inflammation-driven renal diseases that arecurrently treated with systemic steroids can benefit frompeptide-steroid conjugates of the present disclosure that would deliverthe therapeutic specifically to the kidneys at sufficiently highconcentrations to elicit a targeted therapeutic effect, while reducingacute systemic side effects. In patients suffering from chronic disease,this approach can advantageously spare much of the rest of the body fromside effects associated with long-term use of steroidal compounds. Asanother example, the peptide-conjugates of the present disclosure can beused for targeted delivery of iron for kidney preconditioning, thusreducing or preventing toxicity associated with systemic iron delivery.

In some embodiments, a method of treating a condition in a subject inneed thereof comprises administering to the subject a composition orpharmaceutical composition comprising any of the peptides orpeptide-conjugates described herein. For example, in certainembodiments, the composition comprises any of the peptides describedherein. Optionally, the composition comprises a moiety coupled to thepeptide, such as an active agent (e.g., a renal therapeutic agent) orany other moiety described herein. In various embodiments, thepharmaceutical composition comprises any composition of the presentdisclosure or a salt thereof, and any of pharmaceutically acceptablecarriers described herein. In various embodiments, the composition orpharmaceutical composition homes, targets, is directed to, accumulatesin, migrates to, is retained by, or binds to the renal tissue of thesubject following administration. The composition or pharmaceuticalcomposition can provide a therapeutic effect on the renal tissue inorder to treat the condition, as discussed above and herein.

In some embodiments, a method of protecting a kidney of a subject frominjury comprises administering to the subject a composition orpharmaceutical composition comprising any of the peptides orpeptide-conjugates described herein. For example, in certainembodiments, the composition comprises any of the peptides describedherein. Optionally, the composition comprises a moiety coupled to thepeptide, such as an active agent (e.g., a renal therapeutic agent) orany other moiety described herein. In various embodiments, thepharmaceutical composition comprises any composition of the presentdisclosure or a salt thereof, and any of pharmaceutically acceptablecarriers described herein.

In some embodiments, the method further comprises inducing ischemicpreconditioning and/or acquired cytoresistance in the kidney of thesubject. The ischemic preconditioning and/or acquired cytoresistance canprotect the kidney from an injury or insult, as described above andherein. The methods of the present disclosure allow such protectiveresponses to be preemptively induced in order to protect the kidney froman upcoming injury or insult. For example, in certain embodiments, thecomposition or pharmaceutical composition is administered at least 1hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9hours, at least 10 hours, at least 11 hours, at least 12 hours, at least13 hours, at least 14 hours, at least 15 hours, at least 16 hours, atleast 17 hours, at least 18 hours, at least 19 hours, at least 20 hours,at least 21 hours, at least 22 hours, at least 23 hours, at least 24hours, at least 36 hours, at least 48 hours, at least 60 hours, at least72 hours, or at least 96 hours prior to a predicted occurrence of theinjury or insult.

Alternatively or in combination, the present disclosure includes methodsfor inducing a protective response in order to treat an injury or insultthat has already occurred. For example, in certain embodiments, thecomposition or pharmaceutical composition is administered at least 1hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9hours, at least 10 hours, at least 11 hours, at least 12 hours, at least13 hours, at least 14 hours, at least 15 hours, at least 16 hours, atleast 17 hours, at least 18 hours, at least 19 hours, at least 20 hours,at least 21 hours, at least 22 hours, at least 23 hours, at least 24hours, at least 36 hours, at least 48 hours, at least 60 hours, at least72 hours, or at least 96 hours after an occurrence of the injury orinsult.

In some embodiments, the present disclosure provides that any peptide ofthe disclosure including SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, or SEQ ID NO: 451-SEQ ID NO: 569 canas a peptide conjugate with an active agent for treatment of a kidneydisorder. For example, a peptide of SEQ ID NO: 45, SEQ ID NO: 132, orSEQ ID NO: 231 can be conjugated to an active agent and administered toa subject in need thereof to treat a kidney disorder.

In some embodiments, the method further comprises performing a medicalprocedure on the subject. The medical procedure can potentially causeinjury or insult to the subject's kidneys. The method of the presentdisclosure can be used to induce a protective response in order toprotect the kidneys from an injury or insult associated with an upcomingmedical procedure. For example, in certain embodiments, the compositionor the pharmaceutical composition is administered at least 1 hour, atleast 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, atleast 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, atleast 10 hours, at least 11 hours, at least 12 hours, at least 13 hours,at least 14 hours, at least 15 hours, at least 16 hours, at least 17hours, at least 18 hours, at least 19 hours, at least 20 hours, at least21 hours, at least 22 hours, at least 23 hours, at least 24 hours, atleast 36 hours, at least 48 hours, at least 60 hours, at least 72 hours,or at least 96 hours prior to performing the medical procedure.

Alternatively or in combination, the present disclosure includes methodsfor inducing a protective response in order to treat an injury or insultassociated with a medical procedure that has already been performed onthe subject. For example, in certain embodiments, the composition or thepharmaceutical composition is administered at least 1 hour, at least 2hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10hours, at least 11 hours, at least 12 hours, at least 13 hours, at least14 hours, at least 15 hours, at least 16 hours, at least 17 hours, atleast 18 hours, at least 19 hours, at least 20 hours, at least 21 hours,at least 22 hours, at least 23 hours, at least 24 hours, at least 36hours, at least 48 hours, at least 60 hours, at least 72 hours, or atleast 96 hours after performing the medical procedure.

In some embodiments, homing of a peptide of this disclosure to thekidneys can be assessed in an animal model such as those described inZager et al. (Am J Physiol Renal Physiol. 2016 Sep. 1; 311(3):F640-51),Zager et al. (Kidney Int. 2013 October; 84(4):703-12), Zager et al.(Transl Res. 2015 November; 166(5):485-501), Bremlage et al. (BMCNephrol. 2010 Nov. 16; 11:31), Zager et al. (Am J Physiol Renal Physiol.2011 December; 301(6):F1334-45), and Mullins et al. (Dis Model Mech.2016 Dec. 1; 9(12):1419-1433), all of which are incorporated herein byreference.

Multiple peptides or peptide conjugates described herein can beadministered in any order or simultaneously. In some cases, multiplefunctional fragments of peptides derived from toxins or venom, or suchfragments conjugated to active agents, can be administered in any orderor simultaneously. If simultaneously, the multiple peptides or peptideconjugates described herein can be provided in a single, unified form,such as an intravenous injection, or in multiple forms, such assubsequent intravenous dosages.

Peptides, peptide-conjugates, and/or pharmaceutical compositions can bepackaged as a kit. In some embodiments, a kit includes writteninstructions on the use or administration of the peptides,peptide-conjugates, and/or pharmaceutical compositions, in accordancewith the various methods described herein.

EXAMPLES

The following examples are included to further describe some embodimentsof the present disclosure, and should not be used to limit the scope ofthe disclosure.

Example 1 Manufacture of Peptides

The peptide sequence was reverse-translated into DNA, synthesized, andcloned in-frame with siderocalin using standard molecular biologytechniques. (M. R. Green, Joseph Sambrook. Molecular Cloning. 2012 ColdSpring Harbor Press). The resulting construct was packaged into alentivirus, transfected into HEK293 cells, expanded, isolated byimmobilized metal affinity chromatography (IMAC), cleaved with tobaccoetch virus protease, and purified to homogeneity by reverse-phasechromatography. Following purification, each peptide was lyophilized andstored frozen.

Example 2 Radiolabeling of Peptide

This example describes radiolabeling of peptides with standardtechniques. See J Biol Chem. 254(11):4359-65 (1979). The sequences wereengineered to have the amino acids, “G” and “S” at the N terminus. SeeMethods in Enzymology V91:1983 p.570 and Journal of Biological Chemistry254(11):1979 p. 4359. An excess of formaldehyde was used to ensurecomplete methylation (dimethylation of every free amine). The labeledpeptides were isolated via solid-phase extraction on Strata-X columns(Phenomenex 8B-S100-AAK), rinsed with water with 5% methanol, andrecovered in methanol with 2% formic acid. Solvent was subsequentlyremoved in a blowdown evaporator with gentle heat and a stream ofnitrogen gas.

Example 3 Peptide Detectable Agent Conjugates

This example describes the dye labeling of peptides. A peptide of thedisclosure is expressed recombinantly or chemically synthesized, andthen the N-terminus of the peptide is conjugated to an detectable agentvia an NHS ester using DCC or EDC to produce a peptide-detectable agentconjugate. The detectable agent is the fluorophore dye is a cyanine dye,such as Cy5.5 or an Alexa fluorophore, such as Alexa647.

The peptide detectable agent conjugates are administered to a subject.The subject can be a human or a non-human animal. After administration,the peptide detectable agent conjugates accumulate in the kidney. Thesubject, or a biopsy from the subject, can be imaged to visualizelocalization of the peptide detectable agent conjugates to kidney. Insome aspects, visualization of the peptide detectable agent conjugatesin kidney after administration results in diagnosis of kidney damage orany kidney disorder.

Example 4 Peptide Accumulation in Kidneys

This example illustrates peptide accumulation in kidneys in animals withintact kidneys. A peptide of this disclosure was radiolabeled bymethylating lysines and the N-terinus, so the actual binding agentcontained methyl or dimethyl lysine(s) and a methylated or dimethylatedamino terminus. A target dosage of 100 nmol of each peptide carrying10-25 μCi of 14C was administered to Female Harlan athymic nude mice bya tail vein injection. Each peptide was allowed to freely circulatewithin the animal for either 4 hours or 24 hours before the animals areeuthanized and sectioned. Mice were frozen in a hexane/dry ice bath andthen frozen in a block of carboxymethylcellulose. Whole animal sagittalslices were prepared that result in thin frozen sections being availablefor imaging. Thin, frozen sections of animal including imaging oftissues such as brain, tumor, liver, kidney, lung, heart, spleen,pancreas, muscle, adipose, gall bladder, upper gastrointestinal track,lower gastrointestinal track, bone, bone marrow, reproductive track,eye, cartilage, stomach, skin, spinal cord, bladder, salivary gland, andother types of tissues were obtained with a microtome, allowed todesiccate in a freezer, and exposed to phosphoimager plates for aboutten days.

These plates were developed. A signal in tissue darker than the signalexpected from blood in that tissue indicated peptide accumulation in aregion, tissue, structure or cell. High signal in the kidneys indicatedpresence and accumulation of the peptide in the kidneys.

Example 5 Peptide Homing with Therapeutic Agents

This example describes certain exemplary therapeutic agents that areconjugated to a peptide. A peptide of the disclosure is expressedrecombinantly or chemically synthesized and then is conjugated to anexemplary drug, such as paclitaxel or triamcinolone acetonide orbudesonide using techniques known in the art, such as those described inBioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3^(rd)Edition, 2013). One or more drugs is conjugated per peptide, or anaverage of less than one drug is conjugated per peptide.

Coupling of these drugs to a peptide of any of SEQ ID NO: 132, SEQ IDNO: 33; SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, orany of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO:441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 569, or SEQ ID NO: 570targets the drug to the kidney of the subject. One or more drug-peptideconjugates are administered to a human or animal. The resulting peptideor peptide conjugate is administered to a non-human animalsubcutaneously, intravenously, or orally, or is injected directly intokidney intra-articularly. Biodistribution can be assessed by LC/MS,autoradiography, positron emission tomography (PET), or fluorescenceimaging. A peptide or peptide conjugate is homed to kidney.

Any one of these drug-peptide conjugates is used to control pain andinflammation associated with any kidney disorder described herein. Uponadministration and homing of peptide-drug conjugates, the kidney-relatedpain or inflammation condition is alleviated.

Example 6 Peptide Homing to Kidney in Non-Human Animals

This example illustrates a peptide or peptide conjugate of thisdisclosure homing to kidney in non-human animals. Non-human animalsinclude but are not limited to guinea pigs, rabbits, dog, cats, horses,rats, mice, cows, pigs, non-human primates, and other non-human animals.A peptide of the present disclosure is recombinantly expressed orchemically synthesized and is used directly, after radiolabeling, orafter conjugation to a fluorophore or therapeutic compound. The peptideis selected from any one of the peptides of SEQ ID NO: 132, SEQ ID NO:33; SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or anyof SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ IDNO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441,SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 569, or SEQ ID NO: 570. Theresulting peptide or peptide conjugate is administered to a non-humananimal subcutaneously, intravenously, or orally, or is injected directlyinto kidney intra-articularly. Biodistribution is assessed by LC/MS,autoradiography, positron emission tomography (PET), or fluorescenceimaging. A peptide or peptide conjugate is homed to kidney in non-humananimals.

Example 7 Whole Body Fluorescence and Isolated Kidney Fluorescence ofHoming Peptides

This example illustrates whole body fluorescence and isolated kidneyfluorescence of peptide homers of this disclosure. Any peptide of thepresent disclosure is chemically conjugated to one molecule of a nearinfrared fluorophore, at the N-terminus of the peptide via an active NHSester on the dye. A dose of 10 nmol of each peptide conjugated to afluorophore is administered to Female Harlan athymic nude mice, weighing20-25 g, and is administered via tail vein injection. Each experiment isdone at least in duplicate (n=2 mice per group). The peptide fluorophoreconjugate is allowed to freely circulate for the described time periodbefore the mice were euthanized at various time points. Mice areevaluated for peptide distribution of the peptide fluorescence in wholebody imaging and in isolated kidney imaging.

For whole body fluorescence (WBF), at the end of the dosing period, miceare frozen in a hexane/dry ice bath and then embedded in a frozen blockof carboxymethylcellulose. Whole animal sagittal slices are preparedthat result in thin frozen sections for imaging. Thin frozen sectionsare obtained using a microtome and allow visualization of tissues.Sections are allowed to desiccate in a freezer prior to imaging. WBF isperformed on fluorescent sections, which are scanned on a Li-Cor Odysseyscanner at a setting of 169 μm resolution, medium quality, 700 channel,L-2.0 intensity.

For isolated kidney fluorescence studies, mice are euthanized by CO₂asphyxiation at the end of the dosing period. The kidney is removed andimaged on a Spectrum IVIS imager (ex/em: 675 nm. 720 nm) with a 1 secondexposure length and a focal height of 0.5 cm.

Example 8 Whole Body Autoradiography of Homing Peptides

This example illustrates whole body autoradiography of peptide homers ofthis disclosure. Peptides are radiolabeled by methylating lysines at theN-terminus as described in EXAMPLE 2. As such, the peptide may containmethyl or dimethyl lysines and a methylated or dimethylated aminoterminus. A dose of 100 nmol radiolabeled peptide is administered viatail vein injection in Female Harlan athymic nude mice, weighing 20-25g. The experiment is done in at least duplicate (n=2 animals per group).Each radiolabeled peptide is allowed to freely circulate within theanimal for the described time period before the animals were euthanizedand sectioned.

Whole body autoradiography (WBA) sagittal sectioning is performed asfollows. At the end of the dosing period, mice are frozen in ahexane/dry ice bath and then embedded in a frozen block ofcarboxymethylcellulose. Whole animal sagittal slices are prepared thatresult in thin frozen sections for imaging. Thin frozen sections areobtained using a microtome and allow visualization of tissues such asbrain, tumor, liver, kidney, lung, heart, spleen, pancreas, muscle,adipose, gall bladder, upper gastrointestinal tract, lowergastrointestinal tract, bone, bone marrow, reproductive tract, eye,cartilage, stomach, skin, spinal cord, bladder, salivary gland, andmore. Sections are allowed to desiccate in a freezer prior to imaging.

For the autoradiography imaging, tape mounted thin sections are freezedried and radioactive samples are exposed to phosphoimager plates for 7days. These plates are developed and the signal (densitometry) from eachorgan is normalized to the signal found in the cardiac blood of eachanimal. A signal in tissue darker than the signal expected from blood inthat tissue indicates accumulation in a region, tissue, structure, orcell.

Example 9 Peptide Localization in Kidney Extracellular Matrix

This example illustrates localization of peptides of this disclosure inkidney extracellular matrix. In one embodiment, animals are dosed andare processed as described in EXAMPLE 13 and EXAMPLE 14 in animals withintact kidneys. At the end of the dosing period, animals are euthanizedand kidney is optionally removed for use in staining and imagingprocedures. Whole animal sagittal slices are prepared that result inthin frozen sections being available for staining and imaging. Thinfrozen sections or live kidney explants are acquired, stained, andvisualized. Whole animal sagittal slices are prepared that result inthin frozen sections being available for staining and imaging. One ormore kidney components are identified in thin frozen sections or livekidney explants using standard staining techniques: collagen fibrils,glycosaminoglycans, or other aspect of the extracellular matrix. Apeptide of this disclosure is found to localize to a structure in thekidney, localized intracellularly or extracellularly bound, or both.Localization is visualized and confirmed by microscopy.

A peptide of the present disclosure is found to localize to theextracellular matrix in kidney. The peptide is selected from any one ofthe peptides of SEQ ID NO: 132, SEQ ID NO: 33; SEQ ID NO: 4, SEQ ID NO:41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO:451-SEQ ID NO: 529, or SEQ ID NO: 570. The peptide may be bound to oneor more components of the extracellular matrix, such as proteoglycans,glycosaminoglycans, aggrecan, decorin, or collagen, collagen type I, II,III, V, VI, VII and XV, both sulphated and non-sulphatedglycosaminoglycans, glycoproteins and polysaccarides. Localization isvisualized and confirmed by microscopy.

In another embodiment, peptides or peptide-drug conjugates of thisdisclosure are administered in humans and are localized in kidneyextracellular matrix. Kidney fibrosis, focal sclerosis, crescenticglomerulonephritis, and membranoproliferative glomerulonephritis can bediagnosed and treated.

Example 10 Peptide Binding to Kidney Explants

This example illustrates a peptide or peptide conjugation of thisdisclosure homing, targeting, being directed to, migrating to, beingretained by, accumulating in, or binding to human and animal kidneyexplants in culture. The peptide is selected from any one of thepeptides of SEQ ID NO: 132, SEQ ID NO: 33; SEQ ID NO: 4, SEQ ID NO: 41,SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120,SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO:355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQID NO: 529, or SEQ ID NO: 570. Peptides are recombinantly expressed orchemically synthesized and are used directly, after radiolabeling, orafter conjugation to a fluorophore or therapeutic compound. A peptide ofpeptide conjugate of this disclosure is incubated with kidney explantsderived from humans or animals. Peptides of peptide conjugate are foundto bind to kidney explants. The interaction with kidney is confirmedusing various methods that include but are not limited to liquidscintillation counting, confocal microscopy, immunohistochemistry, HPLC,or LC/MS. The peptide shows a higher level of signal than a controlpeptide that is administered that is not a kidney binding peptide.

Example 11 Effects of Peptide on Ion Channels

This example describes the interaction between peptides of the presentdisclosure and ion channels. Ion channels can be associated with painand can be activated in disease states in the kidney, includingvariations in ion channels that cause disease or modulation of ionchannels in order to treat diseases (Kuo et al. Chem Rev. 2012 Dec. 12;112(12):6353-72). A peptide of the disclosure is expressed andadministered in a pharmaceutical composition to a patient to treat akidney condition or disease associated with an ion channel and treatableby binding, blocking, or interacting with the ion channel. Ion channels,such as Nav 1.7, are inhibited by peptides of the present disclosure. Agiven peptide is expressed recombinantly or chemically synthesized,wherein the peptide selected from SEQ ID NO: 132, SEQ ID NO: 33; SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Followingexpression or synthesis, the peptide is used directly or conjugated to atherapeutic compound, such as those described herein. A peptide of thepresent disclosure selectively interacts with ion channels, or ismutated in order to interact with ion channels. For example, a peptideof this disclosure modulates TRPC6 or TRPM6. When the peptide isadministered to a human subject, kidney TRPC6 function is modulated,Ca²⁺ influx is more normalized and focal segmental glomerulosclerosis istreated. Moreover, a peptide of this disclosure is bound to Nav 1.7 by apeptide of this disclosure or Nav 1.7 is blocked by a peptide of thisdisclosure. When the peptide is administered to a human subject, Nav 1.7signaling is reduced in the tissues in or in proximity to the kidney andpain relief is thereby provided.

Example 12 Peptide-Fc Protein Fusions

This example illustrates making and using peptide-Fc protein fusions. Apeptide of SEQ ID NO: 132 was recombinantly expressed with the sequencefor the human IgG1 Fc protein in HEK293 cells to yield a sequence of SEQID NO: 575

(METDTLLLWVLLLWVPGSTGGSGVPINVRCRGSRDCLDPCRRAGMRFGRCINSRCHCTPGGSGGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK)

The sequence of any peptide of this disclosure is expressed as a fusionprotein with either murine or human Fc by adding a secretion signalsequence to the N-terminus and an Fc sequence to the C-terminus. Thiscreates a bivalent molecule with improved secretion properties. Thelarger peptide-Fc fusion is expressed in different mammalian or insectcell lines and is useful as a research reagent and a therapeutic.

Fc fusion to a peptide of SEQ ID NO: 132 to yield a sequence of SEQ IDNO: 575 extends half-life and improves biodistribution of the peptide tothe kidney. Any peptide of this disclosure is co-expressed with Fcprotein to yield Fc-fusion peptides with longer half-life and improvedhoming to kidney. In SEQ ID NO: 575, the secretion signal sequenceMETDTLLLWVLLLWVPGSTG (SEQ ID NO: 576) is followed by the peptide of SEQID NO: 132, and is followed by the sequence for Fc protein. Cleaving canbe imprecise, resulting in cleavage at position 20 or position 21 of SEQID NO: 575.

Example 13 Peptide Conjugate Hydrolysis

This example describes preparation of peptide conjugates having tunablehydrolysis rates. The peptide-drug conjugates described below aresynthesized with the modification that instead of using succinicanhydride, other molecules are used to provide steric hindrance tohydrolysis or an altered local environment at the carbon adjacent to thefinal hydrolyzable ester. In one exemplary conjugate, the peptide-drugconjugate is synthesized with tetramethyl succinic anhydride to generatehindered esters, which causes a decreased rate of hydrolysis. In anotherexemplary conjugate, one methyl group is present at the adjacent carbon.In another exemplary conjugate, two methyl groups are present at theadjacent carbon. In another exemplary conjugate, one ethyl group ispresent at the adjacent carbon. In another exemplary conjugate, twoethyl groups are present at the adjacent carbon. In another exemplaryconjugate, the carbon linker length is increased such as by usingglutaric anhydride instead of succinic anhydride, increasing the localhydrophobicity and lowering the hydrolysis rate. In another exemplaryconjugate, a hydroxyl group is located on the adjacent carbon,increasing the local hydrophilicity and increasing the hydrolysis rate.The rate of hydrolysis in these exemplary conjugates is thereforeadjusted, preventing premature cleavage and ensuring that the majorityof peptide-dexamethasone conjugates accumulate in kidney prior torelease of the drug by hydrolysis but that the dexamethasone is alsoreleased in the kidney in a timely manner.

The resulting peptide conjugates are administered to a human or animalsubcutaneously, intravenously, orally, or injected directly into thekidney to treat disease.

Example 14 Peptide Conjugates with Stable Linkers

This example describes preparation of peptide conjugates with stablelinkers. A peptide of the disclosure (e.g., SEQ ID NO: 132, SEQ ID NO:33, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or anyof SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ IDNO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441,SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570) isexpressed recombinantly or chemically synthesized. The peptide isconjugated to a detectable agent or an active agent via a stable linker,such as an amide linkage or a carbamate linkage. The peptide isconjugated to a detectable agent or an active agent via a stable linker,such as an amide bond using standard1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) ordicylcohexylcarbodiimide (DCC) based chemistry or thionyl chloride orphosphorous chloride-based bioconjugation chemistries.

A peptide and drug conjugated via a linker are described with theformula Peptide-A-B-C-Drug, wherein the linker is A-B-C. A can be astable amide link such as that formed by reacting an amine on thepeptide with a linker containing a tetrafluorophenyl (TFP) ester or anNHS ester. A can also be a stable carbamate linker such as that formedby reacting an amine on the peptide with an imidazole carbamate activeintermediate formed by reaction of CDI with a hydroxyl on the linker. Acan also be a stable secondary amine linkage such as that formed byreductive alkylation of the amine on the peptide with an aldehyde orketone group on the linker. A can also be a stable thioether linkerformed using a maleimide or bromoacetamide in the linker with a thiol inthe peptide, a triazole linker, a stable oxime linker, or a oxacarbolinelinker. B is (—CH2—)_(x)— or a short PEG (—CH₂CH₂O—)_(x) (x is 0-20).Alternatively, spacers within the linker is optional and is included ornot at all. C is an amide bond formed with an amine or a carboxylic acidon the drug, a thioether formed between a maleimide on the linker and asulfhydroyl on the drug, a secondary or tertiary amine, a carbamate, orother stable bonds. Any linker chemistry described in “Current ADCLinker Chemistry,” Jain et al., Pharm Res, 2015 DOI10.1007/s11095-015-1657-7 can be used.

The resulting peptide conjugates are administered to a human or animalsubcutaneously, intravenously, orally, or injected directly into thekidney to treat disease. The peptide is not specifically cleaved fromthe detectable agent or active agent via a targeted mechanism. Thepeptide can be degraded by mechanisms such as catabolism, releasing adrug that is modified or not modified form its native form (Singh,Luisi, and Pak. Pharm Res 32:3541-3571 (2015)). The peptide drugconjugate exerts its pharmacological activity while still intact, orwhile partially or fully degraded, metabolized, or catabolized.

Example 15 Peptide Conjugates with Cleavable Linkers

This example describes preparation of peptide conjugates havingcleavable linkers. The peptide is selected from any one of peptides ofSEQ ID NO: 132, SEQ ID NO: 33, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO:5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO:127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO:529, or SEQ ID NO: 570. A peptide of the disclosure is expressedrecombinantly or chemically synthesized. A peptide and drug areconjugated via a linker and is described with the formulaPeptide-A-B-C-Drug, wherein the linker is A-B-C. A is a stable amidelink such as that formed by reacting an amine on the peptide with alinker containing a tetrafluorophenyl (TFP) ester or an NHS ester. A canalso be a stable carbamate linker such as that formed by reacting anamine on the peptide with an imidazole carbamate active intermediateformed by reaction of CDI with a hydroxyl on the linker. A can also be astable secondary amine linkage such as that formed by reductivealkylation of the amine on the peptide with an aldehyde or ketone groupon the linker. A can also be a stable thioether linker formed using amaleimide or bromoacetamide in the linker with a thiol in the peptide, atriazole linker, a stable oxime linker, or a oxacarboline linker. B is(—CH2—)_(x)— or a short PEG (—CH₂CH₂O—)_(x) (x is 0-20) or other spacersor no spacer. C is an ester bond to the hydroxyl or carboxylic acid onthe drug, or a carbonate, hydrazone, or acylhydrazone, designed forhydrolytic cleavage. The hydrolytic rate of cleavage is varied byvarying the local environment around the ester, including carbon length(—CH2—)_(x), steric hindrance (including adjacent side groups such asmethyl, ethyl, cyclic), hydrophilicity or hydrophobicity. Hydrolysisrate is affected by local pH, such as lower pH in certain compartmentsof the body or of the cell such as endosomes and lysosomes or diseasedtissues. C is a pH sensitive group such as a hydrazone or oxime linkage.Alternatively C is a disulfide bond designed to be released byreduction, such as by glutathione. Alternatively C (or A-B-C) is apeptidic linkage design for cleavable by enzymes. Optionally, aself-immolating group such as pABC is included to cause release of afree unmodified drug upon cleavage (Antibody-Drug Conjugates: Design,Formulation, and Physicochemical Stability, Singh, Luisi, and Pak. PharmRes (2015) 32:3541-3571). The linker is cleaved by enzymes such asesterases, matrix metalloproteinases, cathepsins such as cathepsin B,glucuronidases, a protease, or thrombin. Alternatively, the bonddesigned for cleavage is at A, rather than C, and C could be a stablebond or a cleavable bond. An alternative design is to have stablelinkers (such as amide or carbamate) at A and C and have a cleavablelinker in B, such as a disulfide bond. The rate of reduction ismodulated by local effects such as steric hindrance from methyl or ethylgroups or modulating hydrophobicity/hydrophilicity.

The resulting peptide conjugates are administered to a human or animalsubcutaneously, intravenously, orally, or injected directly into thekidney to treat disease.

Example 16 Acetylsalicylic Acid Peptide Conjugate

This example describes the conjugation of acetylsalicylic acid to apeptide using a lactic acid linker. A conjugate is produced from amixture of (R,S)-acetylsalicylic acid, lactic acid, and a peptide.

The acetylsalicylic acid-lactic acid linker conjugate depicted above isthen reacted with a lysine or the N-terminus of a cystine-dense peptideto create a acetylsalicylic acid-lactic acid-peptide conjugate. Thecystine-dense peptide is selected from the peptides of SEQ ID NO: 132,SEQ ID NO: 33, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO:6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206,SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO:441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570.

Acetylsalicylic acid is currently dosed as an enantiomeric mixture, inwhich enantiomers with a single racemic stereocenter are very difficultto separate. As in the reaction scheme (I), a diastereomer with twochiral centers is created by the addition of a chiral linker such asL-lactic acid. Since diastereomers are easily separated, the activeenantiomer of acetylsalicylic acid conjugated to the lactic acid linkercan be purified prior to conjugation to a peptide. The chemicalsynthesis can use any conjugation techniques known in the art, such asdescribed in Bioconjugate Techniques by Greg Hermanson and in“Ketorolac-dextran conjugates: synthesis, in vitro, and in vivoevaluation:” Acta Pharm. 57 (2007) 441-450, Vyas, Trivedi, andChaturvedi. The conjugate can display anti-inflammatory activity, orfree acetylsalicylic acid is released from the conjugate to provideanti-inflammatory activity. The free acetylsalicylic acid can resultfrom hydrolysis that occurs after administration, such as hydrolysis atthe ester bond. By dosing the conjugate containing the kidney homingpeptide, a higher AUC of acetylsalicylic acid delivery to the kidney maybe achieved than would be achieved by systemic dosing of acetylsalicylicacid alone.

Such peptide-drug conjugates can be made using either a cleavable orstable linker as described herein (e.g., EXAMPLES 14 and 15).

Any one of these drug-peptide conjugates is used to control pain andinflammation associated with any kidney disorder described herein. Uponadministration and homing of peptide-acetylsalicyclic acid conjugates,the kidney-related pain or inflammation condition is alleviated.

Example 17 Acetylsalicylic Acid Peptide Conjugate

This example describes the conjugation of acetylsalicylic acid to apeptide using a PEG linker. A conjugate is produced usingacetylsalicylic acid and a PEG linker, which forms an ester bond thatcan hydrolyze as described in “In vitro and in vivo study ofpoly(ethylene glycol) conjugated ibuprofen to extend the duration ofaction,” Scientia Pharmaceutica, 2011, 79:359-373, Nayak and Jain.Fischer esterification is used to conjugate ibuprofen with a short PEG,e.g., with triethylene glycol, to yield ibuprofen-ester-PEG-OH.

Following preparation of the PEG-ibuprofen conjugate as shown above, thehydroxyl moiety of PEG is activated with N,N′-disuccinimidyl carbonate(DSC) to form ibuprofen-ester-PEG-succinimidyl carbonate, which is thenreacted with a lysine or the N-terminus of a cystine-dense peptide toform an ibuprofen-ester-PEG-peptide conjugate. The cystine-dense peptideis selected from any one of the peptides of sequence SEQ ID NO: 132, SEQID NO: 33, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6,or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206,SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO:441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570.The conjugate can display anti-inflammatory activity, or free ibuprofenis released from the conjugate to provide anti-inflammatory activity.The free ibuprofen can result from hydrolysis that occurs afteradministration, such as hydrolysis at the ester bond.

Ibuprofen-peptide conjugates are administered to a subject in needthereof. The subject can be a human or a non-human animal.

Such peptide-drug conjugates can be made using either a cleavable orstable linker as described herein (e.g., EXAMPLES 14 and 15).

Any one of these drug-peptide conjugates is used to control pain andinflammation associated with any kidney disorder described herein. Uponadministration and homing of peptide-acetylsalicyclic acid conjugates,the kidney-related pain or inflammation condition is alleviated.

Example 18 Dexamethasone Peptide Conjugate

This example describes different methods of conjugating dexamethasonewith a peptide of this disclosure. A peptide of SEQ ID NO: 132, SEQ IDNO: 33, or SEQ ID NO: 196 is recombinantly expressed. Dexamethasone isreadily conjugated to a peptide of this disclosure using a dicarboxylicacid linker. The peptide-dexamethasone conjugate is made by firstconverting dexamethasone to a hemisuccinate by reacting it with succinicanhydride. The hemisuccinate is then converted to a succinate carboxylicacid containing an active ester, using dicyclohexyl carbodiimide (DCC)or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) in the presenceof N-hydroxy succinimide (NETS). This active ester is then reacted witha lysine or the N-terminus of a cystine-dense peptide to create adexamethasone-carboxylic acid-peptide conjugate. Methods such as thosedescribed in “Functionalized derivatives of hyaluronic acidoligosaccharides: drug carriers and novel biomaterials” BioconjugateChemistry 1994, 5, 339-347, Pouyani and Prestwich, and BioconjugateTechniques by Greg Hermanson can be used.

Peptide-dexamethasone conjugates are prepared by coupling dexamethasoneto the peptides of this disclosure using standard coupling-reagentchemistry. For example, dexamethasone conjugates are made by reactingdexamethasone hemigluterate with 1.05 molar equivalents of1,1′-carbonyldiimidazole in anhydrous DMSO in an inert atmosphere. After30 minutes, excess dexamethasone in anhydrous DMSO is added along withtwo molar equivalents of anhydrous trimethylamine. TheN-hydroxysuccinimide ester of the peptide-dexamethasone conjugate isgenerated to form a shelf-stable intermediate for later reaction with anamine-containing carrier. The N-terminal dexamethasone-peptide conjugate(SEQ ID NO: 132B) is verified by electrospray mass spectrometry (ES-MS)within a 10 ppm error.

A peptide of any of the sequences of this disclosure including SEQ IDNO: 132, SEQ ID NO: 33; SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, orSEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, orSEQ ID NO: 570, are conjugated to dexamethasone using the methodsdescribed above.

Such peptide-drug conjugates can be made using either a cleavable orstable linker as described herein (e.g., EXAMPLES 14 and 15).

Example 19 Beclomethasone Monopropionate Peptide Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 45 or SEQID NO: 132 of this disclosure to beclomethasone monopropionate.Beclomethasone monopropionate is readily conjugated to any peptidedisclosed herein via a dicarboxylic acid linker. The dicarboxylic acidlinker is a linear dicarboxylic acid, such as succinic acid, or arelated cyclic anhydride, such as succinic anhydride. Reactions withanhydrides can proceed under simple conditions. For example, thereaction of beclomethasone monopropionate with five molar equivalents ofglutaric anhydride is carried out in anhydrous pyridine at roomtemperature. Reactions with dicarboxylic acids can occur using standardcarbodiimide coupling methods. For example, beclomethasonemonopropionate is reacted with one molar equivalent dimethyl succinicacid, one molar equivalent 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide(or another carbodiimide), and 0.2 molar equivalents of 40-dimethylaminopyridine.

The same methods as described in EXAMPLE 13 are used to adjust the rateof hydrolysis of peptide-beclomethasone monopropionate conjugates,preventing premature cleavage and ensuring that the beclomethasonemonopropionate of peptide-beclomethasone monopropionate conjugatesaccumulate in kidney.

Peptide-beclomethasone monopropionate conjugates are prepared bycoupling beclomethasone monopropionate to the peptides of thisdisclosure using standard coupling-reagent chemistry. Thepeptide-beclomethasone monopropionate conjugate was made by firstconverting beclomethasone monopropionate to a hemisuccinate by reactingit with succinic anhydride. The hemisuccinate was then converted to asuccinate carboxylic acid containing an active ester, using dicyclohexylcarbodiimide (DCC) or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide(EDC) in the presence of N-hydroxy succinimide (NETS). This active esterwas then reacted with a lysine or the N-terminus of a peptide to createa beclomethasone monopropionate-carboxylic acid-peptide conjugate.Methods such as those described in “Functionalized derivatives ofhyaluronic acid oligosaccharides: drug carriers and novel biomaterials”Bioconjugate Chemistry 1994, 5, 339-347, Pouyani and Prestwich, andBioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3^(rd)Edition, 2013) can be used.

Peptide-beclomethasone monopropionate conjugates were prepared bycoupling beclomethasone monopropionate to the peptides of thisdisclosure using standard coupling-reagent chemistry. For example,beclomethasone monopropionate conjugates were made by reactingbeclomethasone monopropionate hemigluterate with 1.05 molar equivalentsof 1,1′-carbonyldiimidazole in anhydrous DMSO in an inert atmosphere.After 30 minutes, excess beclomethasone monopropionate in anhydrous DMSOwas added along with two molar equivalents of anhydrous trimethylamine.The N-hydroxysuccinimide ester of the peptide-beclomethasonemonopropionate conjugate was generated to form a shelf-stableintermediate for later reaction with an amine-containing carrier.

Beclomethasone monopropionate is also readily conjugated to any peptidedisclosed herein via a dicarboxylic acid linker. The dicarboxylic acidlinker is a linear dicarboxylic acid, such as succinic acid, or arelated cyclic anhydride, such as succinic anhydride. Reactions withanhydrides can proceed under simple conditions. For example, thereaction of beclomethasone monopropionate with five molar equivalents ofglutaric anhydride is carried out in anhydrous pyridine at roomtemperature. Reactions with dicarboxylic acids can occur using standardcarbodiimide coupling methods. For example, beclomethasonemonopropionate is reacted with one molar equivalent dimethylsuccinicacid, one molar equivalent 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide(or another carbodiimide), and 0.2 molar equivalents of 40-dimethylaminopyridine. The peptide-beclomethasone monopropionate conjugates areadministered to a subject in need thereof and home, target, are directedto, are retained by, accumulate in, migrate to, and/or bind to kidneys.The subject is a human or animal and has inflammation in the kidneytissues. Upon administration of the peptide-beclomethasonemonopropionate conjugates, the kidney inflammation is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33; SEQID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570.

Such peptide-drug conjugates are made using either a cleavable or stablelinker as described herein (e.g. EXAMPLES 14 and 15).

Example 20 Desciclesonide Peptide Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132, SEQID NO: 33, or SEQ ID NO: 196 of this disclosure to desciclesonide.Ciclesonide is a prodrug that is metabolized in vivo to the activemetabolite desciclesonide. By conjugating desciclesonide to a peptidevia an ester linker, upon hydrolysis the released drug would bedesciclesonide, just as after systemic administration of ciclesonide theactive metabolite desciclesonide is present and active. Desciclesonideis readily conjugated to any peptide disclosed herein via a dicarboxylicacid linker. The dicarboxylic acid linker is a linear dicarboxylic acid,such as succinic acid, or a related cyclic anhydride, such as succinicanhydride. Reactions with anhydrides can proceed under simpleconditions. For example, the reaction of desciclesonide with five molarequivalents of glutaric anhydride is carried out in anhydrous pyridineat room temperature. Reactions with dicarboxylic acids can occur usingstandard carbodiimide coupling methods. For example, desciclesonide isreacted with one molar equivalent dimethylsuccinic acid, one molarequivalent 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (or anothercarbodiimide), and 0.2 molar equivalents of 40-dimethyl amino pyridine.

The same methods as described in EXAMPLE 13 are used to adjust the rateof hydrolysis of peptide-desciclesonide conjugates, preventing prematurecleavage and ensuring that the desciclesonide of peptide-desciclesonideconjugates accumulate in kidney.

Desciclesonide is also readily conjugated to any peptide disclosedherein via a dicarboxylic acid linker. The dicarboxylic acid linker is alinear dicarboxylic acid, such as succinic acid, or a related cyclicanhydride, such as succinic anhydride. Reactions with anhydrides canproceed under simple conditions. For example, the reaction ofdesciclesonide with five molar equivalents of glutaric anhydride iscarried out in anhydrous pyridine at room temperature. Reactions withdicarboxylic acids can occur using standard carbodiimide couplingmethods. For example, desciclesonide is reacted with one molarequivalent dimethylsuccinic acid, one molar equivalent1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (or another carbodiimide),and 0.2 molar equivalents of 40-dimethylamino pyridine

The peptide-desciclesonide conjugates are administered to a subject inneed thereof and home, target, are directed to, are retained by,accumulate in, migrate to, and/or bind to kidneys. The subject is ahuman or animal and has inflammation in the kidney tissues. Uponadministration of the peptide-desciclesonide conjugates, the kidneyinflammation is alleviated.

The peptide-desciclesonide conjugates are administered to a subject inneed thereof and home, target, are directed to, are retained by,accumulate in, migrate to, and/or bind to kidneys. The subject is ahuman or animal and has inflammation in the kidney tissues. Uponadministration of the peptide-desciclesonide conjugates, the kidneyinflammation is alleviated.

The peptide can also be a peptide of SEQ ID NO: 33, SEQ ID NO: 4, SEQ IDNO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO:451-SEQ ID NO: 529, or SEQ ID NO: 570.

Such peptide-drug conjugates are made using either a cleavable or stablelinker as described herein (e.g. EXAMPLES 14 and 15).

Example 21 Peptide-Tofacitinib Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 33 thisdisclosure to Tofacitinib (generic name of Xeljanz). Tofacitinib isreadily conjugated to any peptide disclosed herein via standardchemistries such as those described in, but not limited to, BioconjugateTechniques by Greg Hermanson (Elsevier Inc., 3^(rd) Edition, 2013). Fromone to eight peptides are linked to Xeljanz.

The peptide-tofacitinib conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland is undergoing kidney transplantation. Upon administration and homingof peptide-tofacitinib conjugates, the immune response that could leadto rejection of the kidney transplant is reduced.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 4, SEQID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 22 Peptide-Ustekinumab Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 33 thisdisclosure to ustekinumab. Ustekinumab is readily conjugated to anypeptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) edition, 2013). Alternatively thepeptide-active agent of this Example can be expressed as a fusionprotein. From one to eight peptides are linked to ustekinumab.

The peptide-ustekinumab conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland is undergoing kidney transplantation. Upon administration of thepeptide-ustekinumab conjugates, the immune response that could lead torejection of the kidney transplant is reduced or alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 4, SEQID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 23 Peptide-IL-17 Inhibitor Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 4 thisdisclosure to an IL-17 inhibitor. An IL-17 inhibitor is readilyconjugated to any peptide disclosed herein via standard chemistries suchas those described in, but not limited to, Bioconjugate Techniques byGreg Hermanson (Elsevier Inc., 3^(rd) Edition, 2013).

The peptide-IL-17 inhibitor conjugates are administered to a subject inneed thereof and home, target, are directed to, are retained by,accumulate in, migrate to, and/or bind to kidney. The subject is a humanor animal and has acute kidney injury such as that which can be causedby treatment with cisplatin (Am J Pathol. 2014 May; 184(5):1411-8.Innate IL-17A-producing leukocytes promote acute kidney injury viainflammasome and Toll-like receptor activation). Upon administration andhoming of peptide-IL-17 inhibitor conjugates, the ankylosing spondylitiscondition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 24 Peptide-Iguratimod Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 41 thisdisclosure to iguratimod. Iguratimod is readily conjugated to anypeptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) Edition, 2013).

The peptide-iguratimod conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidneys. The subject is a human or animaland has refractory lupus nephritis. Upon administration and homing ofpeptide-iguratimod conjugates, the refractory lupus nephritis conditionis alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 25 Peptide Mycophenolic Acid Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 5 thisdisclosure to mycophenolic acid. Mycophenolic acid is readily conjugatedto any peptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) Edition, 2013).

The peptide-mycophenolic acid conjugates are administered to a subjectin need thereof and home, target, are directed to, are retained by,accumulate in, migrate to, and/or bind to kidneys. The subject is ahuman or animal and has organ transplantation, infection, cancer, orother kidney disorders. Upon administration and homing ofpeptide-mycophenolic acid conjugates, the organ transplantation,infection, cancer, other kidney disorders condition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 26 Peptide-Tacrolimus Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 6 thisdisclosure to tacrolimus. Tacrolimus is readily conjugated to anypeptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) Edition, 2013).

The peptide-tacrolimus conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidneys. The subject is a human or animaland has organ transplantation, any other kidney disease. Uponadministration and homing of peptide-tacrolimus conjugates, the organtransplantation, any other kidney disease condition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 27 Peptide-Secukinumab Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132, SEQID NO: 4, or SEQ ID NO: 33 to secukinumab. Secukinumab is readilyconjugated to any peptide disclosed herein via standard chemistries suchas those described in, but not limited to, Bioconjugate Techniques byGreg Hermanson (Elsevier Inc., 3^(rd) edition, 2013). From one to eightpeptides are linked to secukinumab. Alternatively the peptide-activeagent of this Example can be expressed as a fusion protein.

The peptide-secukinumab conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has ankylosing spondylitis. Upon administration and homing ofpeptide-secukinumab acid conjugates, the ankylosing spondylitiscondition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 28 Peptide-Sirukumab Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132, SEQID NO: 4, or SEQ ID NO: 33 to sirukumab. Sirukumab is readily conjugatedto any peptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) edition, 2013). From one to eightpeptides are linked to sirukumab. Alternatively the peptide-active agentof this Example can be expressed as a fusion protein.

The peptide-sirukumab conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidneys. The subject is a human or animaland has rheumatoid arthritis, immune diseases of the kidneys. Uponadministration and homing of peptide-sirukumab conjugates, therheumatoid arthritis, immune diseases of the kidneys condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 29 Peptide-Anifrolumab Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132, SEQID NO: 4, or SEQ ID NO: 33 to anifrolumab. Anifrolumab is readilyconjugated to any peptide disclosed herein via standard chemistries suchas those described in, but not limited to, Bioconjugate Techniques byGreg Hermanson (Elsevier Inc., 3^(rd) edition, 2013). From one to eightpeptides are linked to anifrolumab. Alternatively the peptide-activeagent of this Example can be expressed as a fusion protein.

The peptide-anifrolumab conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidneys. The subject is a human or animaland has lupus nephritis. Upon administration and homing ofpeptide-anifrolumab conjugates, the lupus nephritis condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 30 Peptide-Denosumab Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132, SEQID NO: 4, or SEQ ID NO: 33 to denosumab. Denosumab is readily conjugatedto any peptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) edition, 2013). From one to eightpeptides are linked to denosumab. Alternatively the peptide-active agentof this Example can be expressed as a fusion protein.

The peptide-denosumab conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has osteoporosis. Upon administration and homing ofpeptide-denosumab conjugates, the osteoporosis condition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 31 Peptide-Rituximab Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132, SEQID NO: 4, or SEQ ID NO: 33 to rituximab. Rituximab is readily conjugatedto any peptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) edition, 2013). From one to eightpeptides are linked to rituximab. Alternatively the peptide-active agentof this Example can be expressed as a fusion protein.

The peptide-rituximab conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has rheumatoid arthritis, kidney transplant. Upon administration andhoming of peptide-rituximab conjugates, the rheumatoid arthritis, kidneytransplant condition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 32 Peptide-Omalizumab Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132, SEQID NO: 4, or SEQ ID NO: 33 to omalizumab. Omalizumab is readilyconjugated to any peptide disclosed herein via standard chemistries suchas those described in, but not limited to, Bioconjugate Techniques byGreg Hermanson (Elsevier Inc., 3^(rd) edition, 2013). From one to eightpeptides are linked to omalizumab. Alternatively the peptide-activeagent of this Example can be expressed as a fusion protein.

The peptide-omalizumab conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidneys. The subject is a human or animaland has kidney inflammation. Upon administration and homing ofpeptide-omalizumab conjugates, the kidney inflammation condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 33 Peptide-Abatacept Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132 toabatacept. Abatacept is readily conjugated to any peptide disclosedherein via standard chemistries such as those described in, but notlimited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc.,3^(rd) Edition, 2013).

The peptide-abatacept conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidneys. The subject is a human or animaland has lupus nephritis, organ transplant, focal segmentalglomerulosclerosis. Upon administration and homing of peptide-abataceptconjugates, the lupus nephritis, organ transplant, focal segmentalglomerulosclerosis condition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 33, SEQ ID NO: 4, SEQ IDNO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO:451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drug conjugates canbe made using either a cleavable or stable linker as described herein(e.g., EXAMPLES 14 and 15).

Example 34 Peptide-Oxycodone Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 33 tooxycodone. Oxycodone is readily conjugated to any peptide disclosedherein via standard chemistries such as those described in, but notlimited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc.,3^(rd) Edition, 2013).

The peptide-oxycodone conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has kidney-related pain. Upon administration and homing ofpeptide-oxycodone conjugates, the kidney-related pain condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 4, SEQID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 35 Peptide Caspaicin Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 4 tocaspaicin. Caspaicin is readily conjugated to any peptide disclosedherein via standard chemistries such as those described in, but notlimited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc.,3^(rd) Edition, 2013).

The peptide-caspaicin conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has kidney-related pain. Upon administration and homing ofpeptide-caspaicin conjugates, the kidney-related pain condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 36 Peptide-GSK2193874 Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 41 toGSK2193874. GSK2193874 is readily conjugated to any peptide disclosedherein via standard chemistries such as those described in, but notlimited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc.,3^(rd) Edition, 2013).

The peptide-GSK2193874 conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has kidney-related pain. Upon administration and homing ofpeptide-GSK2193874 conjugates, the kidney-related pain condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 37 Peptide BIIB023 Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 5 thisdisclosure to BIIB023. BIIB023 is readily conjugated to any peptidedisclosed herein via standard chemistries such as those described in,but not limited to, Bioconjugate Techniques by Greg Hermanson (ElsevierInc., 3^(rd) Edition, 2013).

The peptide-BIIB023 conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has lupus nephritis. Upon administration and homing ofpeptide-BIIB023 conjugates, the lupus nephritis condition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 38 Peptide-Anakinra Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 6 toanakinra. Anakinra is readily conjugated to any peptide disclosed hereinvia standard chemistries such as those described in, but not limited to,Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., Edition,2013). From one to eight peptides are linked to anakinra. Alternativelythe peptide-active agent of this Example can be expressed as a fusionprotein.

The peptide-anakinra conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has lupus nephritis. Upon administration and homing ofpeptide-anakinra conjugates, the lupus nephritis condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 39 Peptide-IGF-1 Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132 thisdisclosure to IGF-1. IGF-1 is readily conjugated to any peptidedisclosed herein via standard chemistries such as those described in,but not limited to, Bioconjugate Techniques by Greg Hermanson (ElsevierInc., 3^(rd) Edition, 2013). From one to eight peptides are linked toIGF-1. Alternatively the peptide-active agent of this Example can beexpressed as a fusion protein.

The peptide-IGF-1 conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has renal cancer. Upon administration and homing of peptide-IGF-1conjugates, the renal cancer condition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 33, SEQ ID NO: 4, SEQ IDNO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO:451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drug conjugates canbe made using either a cleavable or stable linker as described herein(e.g., EXAMPLES 14 and 15).

Example 40 Peptide-Romosozumab Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132, SEQID NO: 4, or SEQ ID NO: 33 this disclosure to Romosozumab. Romosozumabis readily conjugated to any peptide disclosed herein via standardchemistries such as those described in, but not limited to, BioconjugateTechniques by Greg Hermanson (Elsevier Inc., 3^(rd) edition, 2013). Fromone to eight peptides are linked to romosozumab. Alternatively thepeptide-active agent of this Example can be expressed as a fusionprotein.

The peptide-romosozumab conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has osteoporosis. Upon administration and homing ofpeptide-romosozumab conjugates, the osteoporosis condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 33, SEQ ID NO: 4, SEQ IDNO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO:451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drug conjugates canbe made using either a cleavable or stable linker as described herein(e.g., EXAMPLES 14 and 15).

Example 41 Peptide-ZVAD-fmk Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 33 thisdisclosure to ZVAD-fmk. ZVAD-fmk is readily conjugated to any peptidedisclosed herein via standard chemistries such as those described in,but not limited to, Bioconjugate Techniques by Greg Hermanson (ElsevierInc., 3^(rd) Edition, 2013).

The peptide-ZVAD-fmk conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has acute kidney injury. Upon administration and homing ofpeptide-ZVAD-fmk conjugates, the surgical intervention, surgery foracute kidney injury condition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 4, SEQID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 42 Peptide-S-methylisothiourea Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 4 thisdisclosure to S-methylisothiourea. S-methylisothiourea is readilyconjugated to any peptide disclosed herein via standard chemistries suchas those described in, but not limited to, Bioconjugate Techniques byGreg Hermanson (Elsevier Inc., 3^(rd) Edition, 2013).

The peptide-S-methylisothiourea conjugates are administered to a subjectin need thereof and home, target, are directed to, are retained by,accumulate in, migrate to, and/or bind to kidney. The subject is a humanor animal and has kidney iron overload, renal ischemia reperfusioninjury, or acute kidney injury. Upon administration and homing ofpeptide-S-methylisothiourea conjugates, the kidney iron overload, renalischemia reperfusion injury, or acute kidney injury condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 43 Peptide-P188 Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 41 thisdisclosure to P188. P188 is readily conjugated to any peptide disclosedherein via standard chemistries such as those described in, but notlimited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc.,3^(rd) Edition, 2013).

The peptide-P188 conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has kidney infection or sepsis. Upon administration and homing ofpeptide-P188 conjugates, the kidney infection or sepsis condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 44 Peptide-MIP-3α Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 5 toMIP-3α. MIP-3α is readily conjugated to any peptide disclosed herein viastandard chemistries such as those described in, but not limited to,Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3^(rd)Edition, 2013). From one to eight peptides are linked to MIP-3α.Alternatively the peptide-active agent of this Example is expressed as afusion protein

The peptide-MIP-3α conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to the kidney. The subject is a human oranimal and has kidney injury, repair and regeneration of kidney. Uponadministration and homing of peptide-MIP-3α conjugates, the kidneyinjury, repair and regeneration of kidney condition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 45 Peptide-BMP-7 Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132 toBMP-7. BMP-7 is readily conjugated to any peptide disclosed herein viastandard chemistries such as those described in, but not limited to,Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., 3^(rd)edition, 2013). From one to eight peptides are linked to BMP-7.Alternatively the peptide-active agent of this Example can be expressedas a fusion protein.

The peptide-BMP-7 conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has acute kidney injury or chronic kidney disease. Uponadministration and homing of peptide-BMP-7 conjugates, the kidneycondition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 46 Peptide-Icariin Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 6 toicariin. Icariin is readily conjugated to any peptide disclosed hereinvia standard chemistries such as those described in, but not limited to,Bioconjugate Techniques by Greg Hermanson (Elsevier Inc., Edition,2013).

The peptide-icariin conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidney. The subject is a human or animaland has acute kidney injury or chronic kidney disease. Uponadministration and homing of peptide-icariin conjugates, the kidneycondition is alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 47 Peptide-Captopril Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 132 tocaptopril. Captopril is readily conjugated to any peptide disclosedherein via standard chemistries such as those described in, but notlimited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc.,3^(rd) Edition, 2013).

The peptide-captopril conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidneys. The subject is a human or animaland has diabetic nephropathy. Upon administration and homing ofpeptide-captopril conjugates, the diabetic nephropathy condition isalleviated.

The peptide can also be a peptide of SEQ ID NO: 33, SEQ ID NO: 4, SEQ IDNO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO:451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drug conjugates canbe made using either a cleavable or stable linker as described herein(e.g., EXAMPLES 14 and 15).

Example 48 Peptide-Tofacitinib Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 33 totofacitinib. Tofacitinib is readily conjugated to any peptide disclosedherein via standard chemistries such as those described in, but notlimited to, Bioconjugate Techniques by Greg Hermanson (Elsevier Inc.,3^(rd) Edition, 2013).

The peptide-tofacitinib conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to the kidney. The subject is a human oranimal and has undergone kidney transplant. Upon administration andhoming of peptide-tofacitinib conjugates, immune response to orrejection of the transplanted kidney is reduced or eliminated, and anydamage or injury caused by the kidney transplant is alleviated andregeneration of tissues and/or host acceptance of the transplant ispromoted.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 49 Peptide-Dimethyl Fumarate Conjugates

This example describes conjugation of a peptide of SEQ ID NO: 4 todimethyl fumarate. Dimethyl fumarate is readily conjugated to anypeptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) Edition, 2013). Alternatively,peptide-dimethyl fumarate conjugates can be synthesized by Michaeladdition of a thiol (on the peptide of linker) to dimethyl fumarate asdescribed by Schmidt et al. (Bioorg Med Chem. Jan. 1, 2007;15(1):333-42. Epub Sep. 29, 2006.).

The peptide-dimethyl fumarate conjugates are administered to a subjectin need thereof and home, target, are directed to, are retained by,accumulate in, migrate to, and/or bind to kidneys. The subject is ahuman or animal and has kidney fibrosis. Upon administration and homingof peptide-dimethyl fumarate conjugates, the kidney fibrosis conditionis alleviated.

The peptide can also be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ IDNO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 50 Intra-Kidney Administration of Peptides and PeptideConjugates

This example illustrates direct introduction into kidney byadministration of peptides or peptide conjugates of this disclosure. Apeptide of this disclosure is expressed recombinantly or chemicallysynthesized. In some cases, the peptide is subsequently conjugated to adetectable agent or an active agent. The peptide or peptide conjugate isadministered to a subject in need thereof via administration byinjection or placement directly into the kidney. The kidney ispenetrated by the peptide or peptide conjugate due to the small size ofthe peptide or peptide conjugate, and due to binding of kidneycomponents by the peptide or peptide conjugate. The peptide or peptideconjugate is bound to or retained by the kidney and the residence timein the kidney is longer due to this binding. Optionally, the injectedmaterial is aggregated, is crystallized, or complexes are formed,further extending the depot effect and contributing to longer residencetime.

The peptide can be a peptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 51 Treatment of Gout

This example describes a method for treating gout using peptides of thepresent disclosure. This method is used as a treatment for acute and/orchronic symptoms associated with gout. A peptide of the presentdisclosure is expressed and administered in a pharmaceutical compositionto a patient as a therapeutic for gout. A peptide of the disclosure isrecombinantly or chemically synthesized and then is used directly orconjugated to pegloticase to treat a kidney disorder. A peptide of thedisclosure is recombinantly or chemically synthesized and then is useddirectly or conjugated to probenecid to treat a kidney disorder. Thepeptide is administered in a pharmaceutical composition to a patient andthe peptide is targeted to the kidney affected by gout. One or morepeptides are administered to a human or animal subcutaneously,intravenously, or orally, or is injected directly into the kidney.

The peptide can be a peptide of SEQ ID NO: 41. The peptide can also be apeptide of SEQ ID NO: 132, SEQ ID NO: 33, SEQ ID NO: 4, SEQ ID NO: 41,SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120,SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO:355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQID NO: 529, or SEQ ID NO: 570. Such peptide-drug conjugates can be madeusing either a cleavable or stable linker as described herein (e.g.,EXAMPLES 14 and 15).

Example 52 Treatment or Management of Pain

This example describes a method for treating or managing pain associatedwith a kidney injury or disorder. This method is used as a treatment foracute and/or chronic symptoms associated with a kidney injury ordisorder. A peptide of the disclosure is expressed and administered in apharmaceutical composition to a patient as a therapeutic for pain as aresult of injury or other kidney condition as described herein. Thepeptide of the present disclosure inhibits ion channels, such as Nav1.7. The peptide is expressed recombinantly or chemically synthesized,wherein the peptide selected from SEQ ID NO: 5 or SEQ ID NO: 132, SEQ IDNO: 33, SEQ ID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Alternatively,the peptides of SEQ ID NO: 5 or SEQ ID NO: 132, SEQ ID NO: 33, SEQ IDNO: 4, SEQ ID NO: 41, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO:451-SEQ ID NO: 529, or SEQ ID NO: 570 are mutated to maintain the kidneyhoming function, but to add or increase ion channel inhibition, such asto Nav 1.7. Following expression or synthesis, the peptide is useddirectly or conjugated to a narcotic (e.g. oxycodone), a non-narcoticanalgesic, a counter-irritant (capsaicin), or a pain receptor channelinhibitor (such as the TRPV4 inhibitor GSK2193874). Followingadministration of the peptide, the peptide targets to the kidneyaffected by pain. One or more peptides are administered to a human oranimal subcutaneously, intravenously, or orally, or is injected directlyinto the kidney.

The peptide can be a peptide of SEQ ID NO: 5. The peptide can also be apeptide of SEQ ID NO: 5 or SEQ ID NO: 132, SEQ ID NO: 33, SEQ ID NO: 4,SEQ ID NO: 41, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120,SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO:355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQID NO: 529, or SEQ ID NO: 570. Such peptide-drug conjugates can be madeusing either a cleavable or stable linker as described herein (e.g.,EXAMPLES 14 and 15).

Example 53 Treatment or Management of Pain with Peptides Only

This example describes a method for treating or managing pain associatedwith a kidney injury or disorder. This method is used as a treatment foracute and/or chronic symptoms associated with a kidney injury ordisorder. A peptide of the disclosure is expressed and administered in apharmaceutical composition to a patient as a therapeutic for pain as aresult of injury or other kidney condition as described herein. Thepeptide of the present disclosure inhibits ion channels, such as Nav1.7. The peptide is expressed recombinantly or chemically synthesized,wherein the peptide selected from SEQ ID NO: 6 or SEQ ID NO: 132, SEQ IDNO: 33, SEQ ID NO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Alternatively,the peptides of SEQ ID NO: 6 or SEQ ID NO: 132, SEQ ID NO: 33, SEQ IDNO: 4, SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO:451-SEQ ID NO: 529, or SEQ ID NO: 570 are mutated to maintain the kidneyhoming function, but to add or increase ion channel inhibition, such asto Nav 1.7. Following expression or synthesis, the peptide is useddirectly. Following administration of the peptide, the peptide targetsto the kidney affected by pain. One or more peptides are administered toa human or animal subcutaneously, intravenously, or orally, or isinjected directly into a kidney.

The peptide can be a peptide of SEQ ID NO: 6. The peptide can also be apeptide of SEQ ID NO: 6, SEQ ID NO: 132, SEQ ID NO: 33, SEQ ID NO: 4,SEQ ID NO: 41, or SEQ ID NO: 5, or any of SEQ ID NO: 1-SEQ ID NO: 120,SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO:355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQID NO: 529, or SEQ ID NO: 570.

Example 54 Treatment of Renal Cell Carcinoma

This example illustrates treatment of renal cell carcinoma usingpeptides of the present disclosure. A peptide of the present disclosureis recombinantly expressed or chemically synthesized and are useddirectly, after radiolabeling, or after conjugation to a fluorophore ortherapeutic compound, such as dasatinib. The peptide or peptideconjugate is administered in a pharmaceutical composition to a subjectas a therapeutic for renal cell carcinoma. One or more peptides orpeptide conjugates of the present disclosure are administered to asubject. A subject can be a human or an animal. The pharmaceuticalcomposition is administered subcutaneously, intravenously, orally, orinjected directly into the kidney. The peptides or peptide conjugatestarget kidney affected by renal cell carcinoma.

The peptide can be a peptide of SEQ ID NO: 33. The peptide can also be apeptide of SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, orSEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, orSEQ ID NO: 570. Such peptide-drug conjugates can be made using either acleavable or stable linker as described herein (e.g., EXAMPLES 14 and15).

Example 55 Treatment of Transitional Cell Carcinoma

This example illustrates treatment of transitional cell carcinoma usingpeptides of the present disclosure. A peptide of the present disclosureis recombinantly expressed or chemically synthesized and are useddirectly, after radiolabeling, or after conjugation to a fluorophore ortherapeutic compound, such as dasatinib. The peptide or peptideconjugate is administered in a pharmaceutical composition to a subjectas a therapeutic for transitional cell carcinoma. One or more peptidesor peptide conjugates of the present disclosure are administered to asubject. A subject can be a human or an animal. The pharmaceuticalcomposition is administered subcutaneously, intravenously, orally, orinjected directly into the kidney. The peptides or peptide conjugatestarget kidney affected by transitional cell carcinoma.

The peptide can be a peptide of SEQ ID NO: 33. The peptide can also be apeptide of SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, orSEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, orSEQ ID NO: 570. Such peptide-drug conjugates can be made using either acleavable or stable linker as described herein (e.g., EXAMPLES 14 and15).

Example 56 Treatment for Rapid Pain Relief

This example illustrates rapid pain relief in patients treated forkidney pain with the peptides or peptide conjugates of this disclosure.A peptide of this disclosure is expressed recombinantly or chemicallysynthesized, and then the N-terminus of the peptide is conjugated to anactive agent via an NHS ester to produce a peptide-active agentconjugate. In some aspects, the active agent such as a kidneytherapeutic from TABLE 5 or TABLE 6. In some cases, the peptide alone isadministered to the subject.

The peptide or peptide-active agent conjugate is administered to asubject in need thereof. The subject is a human or non-human animal. Thesubject in need thereof has kidney pain. The peptide or peptideconjugate is delivered via intravenous administration. Uponadministration, the peptide or peptide conjugate rapidly homes tokidney. Rapid pain relief within five minutes to an hour is experiencedby the subject, and pain relief can last as long as over 3 hours.

The peptide can be a peptide of SEQ ID NO: 132. The peptide can also bea peptide of SEQ ID NO: 33, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5,or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO:127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO:529, or SEQ ID NO: 570. Such peptide-drug conjugates can be made usingeither a cleavable or stable linker as described herein (e.g., EXAMPLES14 and 15).

Example 57 Treatment for Lupus Nephritis

This example illustrates treatment of lupus nephritis using peptides orpeptide conjugates of this disclosure. A peptide of the presentdisclosure is recombinantly expressed or chemically synthesized and areused directly, after radiolabeling, or after conjugation to afluorophore or therapeutic compound, such as abatacept or BIIB023.

The peptide or peptide conjugate is administered in a pharmaceuticalcomposition to a subject as a therapeutic for lupus nephritis. Thepeptide is selected from any one of the peptides of SEQ ID NO: 33 or SEQID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6,or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206,SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO:441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570.One or more peptides or peptide conjugates of the present disclosure areadministered to a subject. A subject can be a human or an animal. Thepharmaceutical composition is administered subcutaneously,intravenously, orally, or injected directly. The peptides or peptideconjugates target kidney affected by lupus nephritis.

The peptide can be a peptide of SEQ ID NO: 33. The peptide can also be apeptide of SEQ ID NO: SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ IDNO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355,SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO:529, or SEQ ID NO: 570. Such peptide-drug conjugates can be made usingeither a cleavable or stable linker as described herein (e.g., EXAMPLES14 and 15).

Example 58 Treatment for Acute Kidney Injury (AKI)

This example illustrates treatment of acute kidney injury (AKI) usingpeptides or peptide conjugates of this disclosure. A peptide of thepresent disclosure is recombinantly expressed or chemically synthesizedand are used directly, after radiolabeling, or after conjugation to afluorophore or therapeutic compound, such as such as a kidneytherapeutic from TABLE 5 or TABLE 6.

The peptide or peptide conjugate is administered in a pharmaceuticalcomposition to a subject as a therapeutic for acute kidney injury (AKI).The peptide is selected from any one of the peptides of SEQ ID NO: 132or SEQ ID NO: 33, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ IDNO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO:206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO:570. One or more peptides or peptide conjugates of the presentdisclosure are administered to a subject. A subject can be a human or ananimal. The pharmaceutical composition is administered subcutaneously,intravenously, orally, or injected directly into the kidney. Thepeptides or peptide conjugates target kidney affected by acute kidneyinjury (AKI).

The peptide can be a peptide of SEQ ID NO: 132. The peptide can also bea peptide of SEQ ID NO: SEQ ID NO: 33, SEQ ID NO: 4, SEQ ID NO: 41, SEQID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ IDNO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355,SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO:529, or SEQ ID NO: 570. Such peptide-drug conjugates can be made usingeither a cleavable or stable linker as described herein (e.g., EXAMPLES14 and 15).

Example 59 Treatment for Chronic Kidney Disease (CKD)

This example illustrates treatment of chronic kidney disease (CKD) usingpeptides or peptide conjugates of this disclosure. A peptide of thepresent disclosure is recombinantly expressed or chemically synthesizedand are used directly, after radiolabeling, or after conjugation to afluorophore or therapeutic compound, such as a kidney therapeutic fromTABLE 5 or TABLE 6.

The peptide or peptide conjugate is administered in a pharmaceuticalcomposition to a subject as a therapeutic for chronic kidney disease(CKD). The peptide is selected from any one of the peptides of SEQ IDNO: 33 or SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, orSEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, orSEQ ID NO: 570. One or more peptides or peptide conjugates of thepresent disclosure are administered to a subject. A subject can be ahuman or an animal. The pharmaceutical composition is administeredsubcutaneously, intravenously, orally, or injected directly into thekidney. The peptides or peptide conjugates target kidney affected bychronic kidney disease (CKD).

The peptide can be a peptide of SEQ ID NO: 33. The peptide can also be apeptide of SEQ ID NO: SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ IDNO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355,SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO:529, or SEQ ID NO: 570. Such peptide-drug conjugates can be made usingeither a cleavable or stable linker as described herein (e.g., EXAMPLES14 and 15).

Example 60 Treatment for Hypertensive Kidney Damage

This example illustrates treatment of hypertensive kidney damage usingpeptides or peptide conjugates of this disclosure. A peptide of thepresent disclosure is recombinantly expressed or chemically synthesizedand are used directly, after radiolabeling, or after conjugation to afluorophore or therapeutic compound, such as such as a kidneytherapeutic from TABLE 5 or TABLE 6.

The peptide or peptide conjugate is administered in a pharmaceuticalcomposition to a subject as a therapeutic for hypertensive kidneydamage. The peptide can be a peptide of SEQ ID NO: 33. The peptide isselected from any one of the peptides of SEQ ID NO: 132, SEQ ID NO: 4,SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. One or more peptides orpeptide conjugates of the present disclosure are administered to asubject. A subject can be a human or an animal. The pharmaceuticalcomposition is administered subcutaneously, intravenously, orally, orinjected directly into the kidney. The peptides or peptide conjugatestarget kidney affected by hypertensive kidney damage.

The peptide can be a peptide of SEQ ID NO: 33. The peptide can be apeptide of SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, orSEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, orSEQ ID NO: 570. Such peptide-drug conjugates can be made using either acleavable or stable linker as described herein (e.g., EXAMPLES 14 and15).

Example 61 Treatment for Diabetic Nephropathy

This example illustrates treatment of diabetic nephropathy usingpeptides or peptide conjugates of this disclosure. A peptide of thepresent disclosure is recombinantly expressed or chemically synthesizedand are used directly, after radiolabeling, or after conjugation to afluorophore or therapeutic compound, such as such as a kidneytherapeutic from TABLE 5 or TABLE 6.

The peptide or peptide conjugate is administered in a pharmaceuticalcomposition to a subject as a therapeutic for diabetic nephropathy. Thepeptide is selected from any one of the peptides of SEQ ID NO: 33 or SEQID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6,or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206,SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO:441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570.One or more peptides or peptide conjugates of the present disclosure areadministered to a subject. A subject can be a human or an animal. Thepharmaceutical composition is administered subcutaneously,intravenously, orally, or injected directly into the kidney. Thepeptides or peptide conjugates target kidney affected by diabeticnephropathy.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 62 Treatment for Renal Fibrosis

This example illustrates treatment of renal fibrosis using peptides orpeptide conjugates of this disclosure. A peptide of the presentdisclosure is recombinantly expressed or chemically synthesized and areused directly, after radiolabeling, or after conjugation to afluorophore or therapeutic compound, such as such as a kidneytherapeutic from TABLE 5 or TABLE 6.

The peptide or peptide conjugate is administered in a pharmaceuticalcomposition to a subject as a therapeutic for renal fibrosis. Thepeptide is selected from any one of the peptides of SEQ ID NO: 33 or SEQID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6,or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206,SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO:441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570.One or more peptides or peptide conjugates of the present disclosure areadministered to a subject. A subject can be a human or an animal. Thepharmaceutical composition is administered subcutaneously,intravenously, orally, or injected directly into the kidney. Thepeptides or peptide conjugates target kidney affected by renal fibrosis.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 63 Radiolabeling of Peptide

This example describes radiolabeling of peptides of this disclosure.Several peptides were radiolabeled by reductive methylation with ¹⁴Cformaldehyde and sodium cyanoborohydride with standard techniques. Thesequences were engineered to have the amino acids, “G” and “S” at the Nterminus. See Methods in Enzymology V91:1983 p.570 and JBC 254(11):1979p. 4359. An excess of formaldehyde was used to ensure completemethylation (dimethylation of every free amine). The labeled peptideswere isolated via solid-phase extraction on Strata-X columns (Phenomenex8B-S100-AAK), rinsed with water with 5% methanol, and recovered inmethanol with 2% formic acid. Solvent was subsequently removed in ablowdown evaporator with gentle heat and a stream of nitrogen gas. Thefinal product was verified and characterized by high performance liquidchromatography (HPLC).

Example 64 Accumulation of Peptide in Renal Tissue

This example describes accumulation of peptides of this disclosure inrenal tissue. ¹⁴C-methylated peptides were intravenously dosed into miceat 30-100 nmol per mouse. After 4-24 hours in circulation, deeplyanesthetized mice were euthanized by freezing in dry ice-chilled hexane.Cryosectioning was performed on a Bright-Hacker cryotome, taking 40 μmsagittal sections. Collected sections were allowed to freeze dry at −20°C. for 48-72 hours before being exposed to phosphor imager plates.Plates were exposed for 7 days then scanned on a RayTest CR-Bio35scanner. Analysis was performed with ADA WBA analysis software.

FIGS. 3A and 3B show accumulation of ¹⁴C signal for a peptide of SEQ IDNO: 4 at two time points, 3 hours (FIG. 3A) and 24 hours (FIG. 3B). Thisdata suggests that the peptide is interacting with the kidney, likelycells of the proximal tubule. It is anticipated that freely filteredproteins would not display a persistent signal in the kidneys asobserved here.

Example 65 Engineering of a Peptide for Renal Therapy

This example describes engineering of a peptide of this disclosure forrenal therapy. A selected cystine-dense (e.g., selected from a libraryof over 200,000 identified native cystine-dense peptides), or any one ofSEQ ID NO: 33 or SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO:5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO:127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO:529, or SEQ ID NO: 570, is used as a scaffold for a peptide-basedtherapeutic of the present invention. The peptide is engineered to havetwo functional elements: (1) homing to the specific site of intendedaction in the kidney (e.g., glomerulus, proximal tubule); and (2)therapeutic activity (e.g., block an ion channel, reduce inflammation).The peptide can be engineered to exhibit therapeutic activity in thepresence or the absence of a conjugated therapeutic. The engineering ofthe peptide is accomplished by computational design that replaces nativeamino acids with those selected by computational software or researchersto increase binding and/or activity at the target. Alternatively,mammalian or Pichia display is used, in which many (e.g., tens orhundreds of thousands) of molecules are displayed on cell surfaces, andthose with good binders are selected by flow cytometry. The leadingcandidates (e.g., identified by deep sequencing of flow-captured cells)are then used as the basis for further design. Iterative rounds ofevolution using the above and related techniques are used to discoverpeptides that have both kidney targeting and therapeutic activity in theabsence of a “payload” conjugate. The peptides are used in a renaltherapy or renal therapeutic application of the present disclosure.

Example 66 Peptide Immunogenicity

This example illustrates the testing of the immunogenicity of a peptide.NetMHC II version 2.3 prediction software is used to identifyimmunogenic peptides based on a neural network alignment algorithm thatpredicts peptide binding to MHC Class II molecules.

The NetMHC II prediction software is utilized to determine the putativepeptide binding capability to DR, DQ, and DP MHC II alleles and thestrength of the interaction between peptide and MHC II molecules. Usingsuch methods identifies the resulting immunogenicity score of selectpeptides. The numbers of strong versus weak peptides are tallied intoeach major MHC allele group (DR, DQ, and DP). Additionally, the numbersof ‘unique strong’ and ‘unique weak core’ peptides are also tallied.These data are used to predict which peptides are less likely to inducean immunogenic response in patients. For example, the stronger a peptidebinds to an allele, the more likely it is to be presented in aMHC/peptide combination on an antigen presenting cell, thus triggeringan immune response, and a peptide that is predicted to bind to feweralleles is more likely to have weaker binding to given alleles andshould be less immunogenic.

Example 67 Peptide-Budesonide Conjugate

This example describes conjugation of a peptide of any one of SEQ ID NO:33 or SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ IDNO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, orSEQ ID NO: 570 to budesonide. Budesonide is readily conjugated to anypeptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) edition, 2013) or by any of the methodsdescribed in the preceding EXAMPLES.

The peptide-budesonide conjugates are administered to a subject in needthereof and home, target, are directed to, are retained by, accumulatein, migrate to, and/or bind to kidneys. The subject is a human or animaland has inflammation in the kidney tissues. Upon administration andhoming of peptide-budesonide conjugates, the inflammation in the kidneytissues is alleviated.

Example 68 Peptide-Dexamethasone Conjugate

This example describes conjugation of a peptide of any one of SEQ ID NO:33 or SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ IDNO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, orSEQ ID NO: 570 to dexamethasone. Dexamethasone is readily conjugated toany peptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) edition, 2013) or by any of the methodsdescribed in the preceding EXAMPLES.

The peptide-dexamethasone conjugates are administered to a subject inneed thereof and home, target, are directed to, are retained by,accumulate in, migrate to, and/or bind to kidneys. The subject is ahuman or animal and has inflammation in kidney tissues. Uponadministration and homing of peptide-dexamethasone conjugates, theinflammation in kidney tissues is alleviated.

Example 69 Peptide-Triamcinalone Acetonide Conjugate

This example describes conjugation of a peptide of any one of SEQ ID NO:33 or SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5 or SEQID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ IDNO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO:362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, orSEQ ID NO: 570 to triamicinalone acetonide. Triamicinalone acetonide isreadily conjugated to any peptide disclosed herein via standardchemistries such as those described in, but not limited to, BioconjugateTechniques by Greg Hermanson (Elsevier Inc., 3^(rd) edition, 2013 or byany of the methods described in the preceding EXAMPLES.

The peptide-triamicinalone acetonide conjugates are administered to asubject in need thereof and home, target, are directed to, are retainedby, accumulate in, migrate to, and/or bind to kidneys. The subject is ahuman or animal and has inflammation in kidney tissues. Uponadministration and homing of peptide-triamicinalone acetonideconjugates, the inflammation in kidney tissues is alleviated.

Example 70 Peptide-Desciclesonide Acetonide Conjugate

This example describes conjugation of a peptide of any one of SEQ ID NO:33 or SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, SEQ IDNO: 6 or SEQ ID NO: 1-SEQ ID NO: 529, or SEQ ID NO: 570 todesciclesonide acetonide. Desciclesonide acetonide is readily conjugatedto any peptide disclosed herein via standard chemistries such as thosedescribed in, but not limited to, Bioconjugate Techniques by GregHermanson (Elsevier Inc., 3^(rd) edition, 2013) or by any of the methodsdescribed in the preceding EXAMPLES.

The peptide-desciclesonide acetonide conjugates are administered to asubject in need thereof and home, target, are directed to, are retainedby, accumulate in, migrate to, and/or bind to kidneys. The subject is ahuman or animal and has inflammation in kidney tissues. Uponadministration and homing of peptide-desciclesonide acetonideconjugates, the inflammation in kidney tissues is alleviated.

Example 71 Method of Peptide Synthesis

This example describes the synthesis of any one of SEQ ID NO: 33 or SEQID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6,or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206,SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO:441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570.

A selected peptide is made using Solid Phase Peptide Synthesis (SPPS).After release of the peptide from the solid phase, the peptide waspurified prior to folding by oxidation in solution. The folded peptidewas further purified by reversed-phase chromatography and lyophilized asa TFA salt. The final peptide product has a purity of greater than 90%,greater than 95%, greater than 98%, about 95-96%, and a mass in Da thatis the estimated molecular mass of the selected peptide which confirmsits identity as the selected peptide.

Example 72 Treatment of a Kidney Condition with a Peptide of theDisclosure

This example describes treatment of a kidney condition with peptides ofthis disclosure. A peptide of the disclosure (e.g., any of the peptidesof SEQ ID NO: 33 or SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ IDNO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ IDNO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355,SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO:529, or SEQ ID NO: 570) is expressed recombinantly or chemicallysynthesized. The peptide is administered to a human or animal, where itbinds to renal tissue and exhibits a therapeutic effect, e.g., viaantioxidant or anti-inflammatory actions. For example, a peptide of thepresent disclosure is taken up by the proximal tubules, and gains accessto and suppresses intracellular injury pathways. As another example, apeptide of the present disclosure migrates to the renal interstitium andinhibits interstitial inflammation and prevents renal fibrosis.

Example 73 Treatment of a Kidney Condition with a Peptide-Conjugate ofthe Disclosure

This example describes treatment of a kidney condition with apeptide-conjugation of this disclosure. A peptide of the disclosure(e.g., any of the peptides of SEQ ID NO: 33 or SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570) is expressedrecombinantly or chemically synthesized. The peptide is then conjugatedto a therapeutic agent, such as deferoxamine, dexamethasone, or anotheranti-inflammatory agent, a chemotherapeutic, or a steroid. Coupling ofthe therapeutic agent to the peptide targets the therapeutic agent tothe kidney. One or more peptide-conjugates are administered to a humanor animal. The therapeutic agent is presented in the kidney at adequateconcentration to provide a therapeutic effect, such as an antioxidant,anti-inflammatory, or a chemotherapeutic effect. Optionally, theconcentration of the therapeutic agent in other tissues is sufficientlylow so to cause few or no undesirable side effects.

For example, a peptide of the present disclosure conjugated todexamethasone or other potent anti-inflammatory agents is used astherapy for lupus affecting the kidney, vasculitis, Goodpasture'sdisease, focal segmental glomerulosclerosis, nephritic syndrome, orother renal disorders caused by inflammatory processes.

As another example, a peptide of the present disclosure is used todeliver a chemotherapeutic for treating renal cell carcinoma.

In a further example, a peptide of the present disclosure is used todeliver steroids for treating polycystic renal disease.

Example 74 Eliciting a Protective Response in the Kidney with a Peptideof the Disclosure

This peptide describes eliciting a protective response in the kidneywith peptides of this disclosure. A peptide of the disclosure (e.g., anyof the peptides of SEQ ID NO: 33 or SEQ ID NO: 132, SEQ ID NO: 4, SEQ IDNO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO:451-SEQ ID NO: 529, or SEQ ID NO: 570) is expressed recombinantly orchemically synthesized. The peptide is administered to a human oranimal, where it binds to renal tissue and induces ischemicpreconditioning or acquired cytoresistance in the kidney. The peptide isadministered to the subject prior to an anticipated injury to thekidney, such as surgery or imaging. The injury that occurs to the kidneyis reduced by the peptide. Optionally, the progression of acute kidneyinjury to chronic kidney disease is reduced by the protective response.

Example 75 Protecting the Kidney from Nephrotoxic Agents with a Peptideof the Disclosure

This example describes protecting the kidney from nephrotoxic agentswith peptides of this disclosure. A peptide of the disclosure (e.g., anyof the peptides of SEQ ID NO: 33 or SEQ ID NO: 132, SEQ ID NO: 4, SEQ IDNO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQ ID NO:120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO: 216-SEQID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQ ID NO:451-SEQ ID NO: 529, or SEQ ID NO: 570) is expressed recombinantly orchemically synthesized. The peptide is administered to a human oranimal, where it binds to renal tissue, e.g., at megalin-cubulin bindingsites. The peptide is administered to the subject prior to or currentlywith a nephrotoxic agent (e.g., aminoglycoside antibiotics such asgentamicin, vancomycin, and minocycline, chemotherapeutics such ascisplatin, immunoglobulins, mannitol, NSAIDs, cyclosporin,cyclophosphamide, radiocontrast dyes) in order to minimize its damagingeffects, e.g., by blocking megalin-cubulin binding sites so that thenephrotoxic agent passes through the kidneys.

Example 76 Eliciting a Protective Response in the Kidney with aPeptide-Conjugate of the Disclosure

This example describes eliciting a protective response in the kidneywith a peptide-conjugation of this disclosure. A peptide of thedisclosure (e.g., any of the peptides of SEQ ID NO: 33 or SEQ ID NO:132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or anyof SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ IDNO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441,SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570) isexpressed recombinantly or chemically synthesized. The peptide is thenconjugated to a renal protective agent, such as a deferoxamine, or achelate or porphyrin complex (e.g., hemin, an EDTA-Fe complex). Couplingof the protective agent to the peptide targets the protective agent toappropriate regions of the kidney with a suitable pharmacokineticprofile. One or more peptide-conjugates are administered to a human oranimal. The peptide conjugate is administered to the subject prior to ananticipated injury to the kidney, such as surgery or imaging. The renaltissue injury that occurs in the kidney is reduced by the peptideconjugate. Optionally, the progression of kidney injury to chronickidney disease is reduced by the protective response.

For example, a peptide of the present disclosure is conjugated to hemin,which signals through the heat shock/heme reactive element pathway. Onceintracellular localization is achieved, an upregulation of a set ofdiverse cytoprotective proteins occurs. The peptide-hemin conjugate isadministered to a subject who will undergo high-risk surgeries orradiocontrast administration. The peptide-hemin conjugate isadministered one day prior to the procedure in order to allow sufficienttime for the upregulation of protective proteins to occur.

As another example, a peptide of the present disclosure is used todeliver iron to the kidney, either as a chelate or porphyrin complex, inorder to alter gene expression profiles and induce expression ofcytoprotective proteins.

Example 77 Confocal Imaging of Kidneys

This example illustrates confocal imaging of kidneys from miceadministered peptides of the present disclosure. A dose of 10 nmol ofAlexFluor 647 (AF647) labeled peptide was administered intravenously inmice (2 per group). Mice were euthanized 20 hours post-peptideadministration and kidneys were harvested and cut into 2 mm sections.Adjacent sections were scanned on an Odyssey instrument at 54 μmresolution in the 700 nm channel or imaged on a Zeiss laser scanningmicroscope (LSM) 780 confocal microscope at 6× and 20× magnification.

FIG. 5 shows fluorescence of kidney sections from mice, in which eachmouse received 10 nmol free fluorophore (AF647), 10 nmol SEQ ID NO: 41conjugated to AF647, 10 nmol SEQ ID NO: 5 conjugated to AF647, or 10nmol SEQ ID NO: 33 conjugated to AF647. Each kidney was from anindependent mouse (2 mice per group).

FIG. 6 shows SEQ ID NO: 5 conjugated to AF647 and SEQ ID NO: 41conjugated to AF647 fluorescence signal in confocal images of the kidneycortex. FIG. 6A shows fluorescence signal of SEQ ID NO: 5 conjugated toAF647 in the kidney cortex 20 hours after of administration of 10 nmolof the peptide-dye conjugate at 6× magnification. FIG. 6B showsfluorescence signal of SEQ ID NO: 5 conjugated to AF647 in the kidneycortex 20 hours after administration of 10 nmol of the peptide-dyeconjugate at 20× magnification. FIG. 6C shows fluorescence signal of SEQID NO: 5 conjugated to AF647 in the kidney cortex 20 hours afteradministration of 10 nmol of the peptide-dye conjugate at 6×magnification. FIG. 6D shows fluorescence signal of SEQ ID NO: 5conjugated to AF647 in the kidney cortex 20 hours after ofadministration of 10 nmol of the peptide-dye conjugate at 20×magnification.

FIG. 7 shows SEQ ID NO: 33 conjugated to AF647 fluorescence signal inconfocal images of the kidney cortex. FIG. 7A shows fluorescence signalof SEQ ID NO: 33 conjugated to AF647 in the kidney cortex 20 hours afteradministration of 10 nmol of the peptide-dye conjugate at 6×magnification. FIG. 7B shows fluorescence signal of SEQ ID NO: 33conjugated to AF647 in the kidney cortex 20 hours after administrationof 10 nmol of the peptide-dye conjugate at 20× magnification. FIG. 7Cshows fluorescence signal in the kidney cortex 20 hours afteradministration of 10 nmol of a lysozyme-dye conjugate at 6×magnification. FIG. 7D shows fluorescence signal in the kidney cortex 20hours after of administration of 10 nmol of a lysozyme-dye conjugate at20× magnification.

Therefore, FIG. 5 shows that the peptides can accumulate the conjugateddye in the cortex of the kidney, and FIG. 6 and FIG. 7 show that thepeptides can accumulate the conjugate dye in the proximal tubules in thekidney, as confirmed by the positive control lysozyme which has beenshown to accumulate in the proximal tubules.

Example 78 Competitive Renal Uptake Studies

This example describes competitive uptake studies of peptides of thisdisclosure in kidneys. Peptides of this disclosure were compared toknown kidney homers (“competitors”) to assess the efficiency andstrength of kidney targeting. Three competitors were tested against apeptide of SEQ ID NO: 4, and kidney uptake was quantified byfluorescence imaging of whole organs on a Spectrum IVIS imager.

FIG. 8 shows competitive renal uptake between a peptide of SEQ ID NO: 4conjugated to AlexaFluor647 (AF647) and an unlabeled SEQ ID NO: 4peptide 4 hours after intravenous administration of 2 nmol of SEQ ID NO:4-AF647 co-injected with either 0 nmol of SEQ ID NO: 4 peptide (“lowAF”), 10 nmol of SEQ ID NO: 4 co-injected with 2 nmol of SEQ ID NO:4-AF647 (5:1), or 50 nmol of SEQ ID NO: 4 co-injected with 2 nmol of SEQID NO: 4-AF647 (25:1). Kidneys from uninjected mice were used as anegative control. Fluorescence signal in each group was quantified todetermine the average radiant efficiency in the kidneys from three miceper cohort. Data are shown as mean and error bars indicate standarddeviation. A p-value of 0.0081 was calculated by a T-test, and the errorbars indicate standard deviation. In this experiment, the unlabeled SEQID NO: 4 peptide competed with the SEQ ID NO: 4-AF647 as shown bydecreased fluorescence and thus, decreased accumulation of the dyelabeled peptide in the kidney. This indicates that SEQ ID NO: 4 peptideuptake was specific and saturable. In contrast, FIG. 11 shows nocompetitive renal uptake between a peptide of SEQ ID NO: 4 conjugated toAlexaFluor647 (AF647) and unlabeled KKEEEKKEEEKKEEEKK peptide (SEQ IDNO: 571, a known renal targeting peptide; see Bioconjug Chem. 2016 Apr.20; 27(4):1050-7) 1 hour after intravenous administration of 2 nmol of apeptide of SEQ ID NO: 4-AF647, 2 nmol of a peptide of SEQ ID NO: 4-AF647co-injected with 100 nmol of an unlabeled peptide of SEQ ID NO: 571(1:50), or 2 nmol of peptide of SEQ ID NO: 4-AF647 co-injected with 2000nmol of an unlabeled peptide of SEQ ID NO: 571 (1:1000). Fluorescencesignal in each group was quantified to determine the average radiantefficiency in the kidneys from three mice per cohort. Data are shown asmean and error bars indicate standard deviation. Kidney uptake of apeptide of SEQ ID NO: 4-AF647 was not dampened by SEQ ID NO: 571 peptideeven at the highest ratio of competitor. The SEQ ID NO: 571 peptidefailed to compete with uptake of the peptide of SEQ ID NO: 4 in kidneys.Since SEQ ID NO: 571 has been hypothesized to bind to megalin, theseresults potentially indicate that SEQ ID NO: 4 peptide may accumulate inthe proximal tubules by a different mechanism or receptor, or may bindto megalin more strongly than SEQ ID NO: 571 peptide. FIG. 22 also showsno competitive renal uptake between a peptide of SEQ ID NO: 4 conjugatedto AlexaFluor647 (AF647) and a control peptide conjugated to AF647(control peptide-AF647), 4 hours after intravenous administration of 10nmol of a peptide of SEQ ID NO: 4-AF647 or 10 nmol of controlpeptide-AF647. Fluorescence signal in each group was quantified todetermine the average radiant efficiency in the kidneys from three miceper cohort. Data are shown as mean and error bars indicate standarddeviation. A p-value of 0.015 was calculated by a Student's unpairedt-test. The peptide of SEQ ID NO: 4 was taken up in the kidneys to asignificantly higher extent than the control peptide.

Example 79 Preclinical Testing of Competitive Inhibition of ToxicProtein Uptake by Kidneys

This example illustrates preclinical validation in mice of competitiveinhibition of toxic protein uptake by kidneys. Myoglobin is a toxicprotein, which can accumulate in proximal tubules via megalin-mediatedendocytosis. Peptides of this disclosure, which are injected in asubject at the time of kidney myoglobin exposure, will compete formegalin-mediated uptake.

A subject is injected intramuscularly with glycerol, leading to muscleinjury with myoglobin release (also referred to herein as a “myoglobinchallenge”). The subject in preclinical testing is a mouse. At the timeof myoglobin injection, the subject is intravenously administered apeptide of this disclosure at one of a range of doses (0.1-2 mg/mouse)or saline as a negative control. Four hours after administration, thedegree of myoglobin uptake by the kidney is tested using aspectrophotometric assay. The severity of myoglobin injury is assessedby testing for siderocalin mRNA (a biomarker of this process)upregulation.

Increasing the dose of the administered peptide of this disclosurecauses a reciprocal decrease in myoglobin uptake in the kidney.Treatment of a subject with peptides of this disclosure results indose-dependent blunting of siderocalin mRNA induction. In negativecontrol subjects, which do not receive a peptide of this disclosure,glycerol injection causes an approximate 10-fold increase in siderocalinmRNA expression.

Example 80 Preclinical Testing of Alleviation of Renal Inflammation

This example illustrates preclinical validation in a subject of thealleviation of renal inflammation following endotoxin injection. Apeptide of the present disclosure is conjugated to dexamethasone asdescribed for desciclesonide peptide conjugates in EXAMPLE 18. Thesubject in preclinical testing is a mouse. Mice are injectedintravenously with E. Coli endotoxin at 1 mg/kg to induce renalinflammation and co-injected intravenously either with saline as anegative control or with increasing doses of a peptide of thisdisclosure (0.1-2 mg/mouse). Four hours post-administration, severity ofrenal inflammation is assessed by measuring inflammatory mediator mRNAs,such as TNFα and monocyte chemoattractant protein (MCP)-1.

Co-injection of peptides of this disclosure causes dose-dependentblunting of mRNA upregulation. In negative control subjects, which donot receive a peptide of this disclosure, endotoxin injections inducesan approximate 5-fold increase in TNFα and MCP-1 mRNA expression within4 hours of endotoxin injection.

Example 81 Peptide Detectable Agent Conjugates

This example describes the dye labeling of peptides. A peptide of thedisclosure is expressed recombinantly or chemically synthesized, andthen the N-terminus of the peptide is conjugated to an detectable agentvia an NHS ester using DCC or EDC to produce a peptide-detectable agentconjugate. The detectable agent is the fluorophore dye is a cyanine dye,such as Cy5.5 or an Alexa fluorophore, such as Alexa647.

The peptide detectable agent conjugates are administered to a subject.The subject can be a human or a non-human animal. After administration,the peptide detectable agent conjugates home to the kidneys. Thesubject, or a biopsy from the subject, is imaged to visualizelocalization of the peptide detectable agent conjugates to the kidney.In some aspects, diagnosis of renal disorders is based on thevisualization of the peptide detectable agent conjugates in kidneysafter administration.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 82 Peptide Deferasirox Conjugates

This example describes conjugation of peptides of this disclosure todeferasirox, an iron chelator. A peptide of the disclosure is expressedrecombinantly or chemically synthesized, and then the N-terminus of thepeptide is conjugated to deferasiroxvia an NHS ester using DCC or EDC toproduce a peptide-deferasiroxconjugate. Alternatively, a peptide can beconjugated to a deferasiroxby common techniques known in the art, suchthose described in Bioconjugate Techniques by Greg T. Hermanson(Elsevier Inc., 3^(rd) Edition, 2013).

The peptide-deferasiroxconjugates are administered to a subject. Thesubject can be a human or non-human animal. The subject can have apre-existing condition, such as iron poisoning. After administration,the peptide-deferasiroxconjugates home to the kidneys.Peptide-deferasiroxconjugates are used to treat iron poising byenhancing elimination of iron in urine.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 83 Peptide Olmesartan Conjugates

This example describes conjugation of peptides of this disclosure toolmesartan. A peptide of the disclosure is expressed recombinantly orchemically synthesized, and then the N-terminus of the peptide isconjugated to olmesartanto produce a peptide-olmesartanconjugate.Optionally, a hydrolytically labile ester linkage is used in theconjugation, such that free olmesartanis released after delivery to thekidney and/or proximal tubule.

The peptide-olmesartanconjugates are administered to a subject. Thesubject can be a human or non-human animal. Optionally, a higher ratioof olmesartanis seen in the kidney versus in serum after administrationof the peptide-olmesartanconjugate than when olmesartanis administeredalone. The subject can have a pre-existing condition, such as a renaldisease. After administration, the peptide-olmesartanconjugates home tothe kidneys. Peptide-olmesartanconjugates is used to treat patients withrenal disease.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 84 Peptide Poly-L-Glutamic Acid Polymer Conjugates

This example describes conjugation of peptides of this disclosure topoly-L-glutamic acid polymers. A peptide of the disclosure is expressedrecombinantly or chemically synthesized, and then the N-terminus of thepeptide is conjugated to poly-L-glutamic acid polymers to produce apeptide-poly-L-glutamic acid polymer conjugate.

The peptide-poly-L-glutamic acid polymers conjugates are administered toa subject. The subject can be a human or non-human animal. The subjectcan have a pre-existing condition, such as kidney disease. Afteradministration, the peptide-poly-L-glutamic acid polymers conjugates arehomed to the kidneys. Peptide-poly-L-glutamic acid polymer conjugatesare used to prevent loss in kidney function in a subject.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 85 Peptide Tirilazad Conjugates

This example describes conjugation of peptides of this disclosure toTirilazad. A peptide of the disclosure is expressed recombinantly orchemically synthesized, and then the N-terminus of the peptide isconjugated to an Tirilazad to produce a peptide-Tirilazad conjugate. TheTirilazad can be glutathione or N acetyl cysteine.

The peptide-Tirilazad conjugates are administered to a subject. Thesubject can be a human or non-human animal. The subject can have apre-existing condition, such as diabetic nephropathy or post-ischemic ornephrotoxic AKI. After administration, the peptide-Tirilazadconjugatesare homed to the kidneys. Peptide-Tirilazadconjugates are used toprevent loss in kidney function and protect renal function in subjectswith one of the above pre-existing conditions.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 86 Prophylaxis Against Acute Kidney Injury

This example describes prophylaxis against acute kidney injury (AKI)with the peptides of the present disclosure. A peptide of thisdisclosure is expressed recombinantly or chemically synthesized. In somecases, the peptide is subsequently conjugated to an active agent. Thepeptide or peptide-active agent conjugate is administered to a subjectin need thereof. The subject is a human or non-human animal. The subjectin need thereof is at risk for acute kidney injury as a result ofcardiovascular surgery, radiocontrast nephropathy, orcisplatin/carboplatin use. The peptide or peptide-conjugate is deliveredvia intravenous administration. Upon administration, the peptide orpeptide conjugate rapidly targets the kidneys, and is used asprophylaxis against AKI.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 87 Treatment of Established Acute Kidney Injury

This example describes treatment of acute kidney injury (AKI) with thepeptides of the present disclosure. A peptide of this disclosure isexpressed recombinantly or chemically synthesized. In some cases, thepeptide is subsequently conjugated to an active agent. The peptide orpeptide-active agent conjugate is administered to a subject in needthereof. The subject is a human or non-human animal. The subject in needthereof has ischemic renal injury, endotoxemia-induced AKI, orestablished nephrotoxic AKI. The peptide or peptide-conjugate isdelivered via intravenous administration. Upon administration, thepeptide or peptide conjugate rapidly targets the kidneys, and is used totreat AKI.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 88 Treatment of Diabetic Nephropathy

This example describes treatment of diabetic nephropathy with thepeptides of the present disclosure. A peptide of this disclosure isexpressed recombinantly or chemically synthesized. In some cases, thepeptide is subsequently conjugated to an active agent. The peptide orpeptide-active agent conjugate is administered to a subject in needthereof. The subject is a human or non-human animal. The subject in needthereof is diagnosed with diabetic nephropathy. The peptide orpeptide-conjugate is delivered via intravenous administration. Uponadministration, the peptide or peptide conjugate rapidly targets thekidneys, and is used to treat diabetic nephropathy.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 89 Treatment of Hypertensive Nephrosclerosis

This example describes treatment of hypertensive nephrosclerosis withthe peptides of the present disclosure. A peptide of this disclosure isexpressed recombinantly or chemically synthesized. In some cases, thepeptide is subsequently conjugated to an active agent. The peptide orpeptide-active agent conjugate is administered to a subject in needthereof. The subject is a human or non-human animal. The subject in needthereof is has hypertensive nephrosclerosis. The peptide orpeptide-conjugate is delivered via intravenous administration. Uponadministration, the peptide or peptide conjugate is rapidly targeted tothe kidneys, and is used to treat hypertensive nephrosclerosis.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 90 Treatment of Chronic Glomerulonephritis

This example describes treatment of chronic glomerulonephritis with thepeptides of the present disclosure. A peptide of this disclosure isexpressed recombinantly or chemically synthesized. In some cases, thepeptide is subsequently conjugated to an active agent. The peptide orpeptide-active agent conjugate is administered to a subject in needthereof. The subject is a human or non-human animal. The subject in needthereof is diagnosed with idiopathic or secondary chronicglomerulonephritis. The peptide or peptide-conjugate is delivered viaintravenous administration. Upon administration, the peptide or peptideconjugate rapidly targets the kidneys, and is used to treat chronicglomerulonephritis.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 91 Treatment of Hereditary Nephropathy

This example describes treatment of hereditary nephropathy with thepeptides of the present disclosure. A peptide of this disclosure isexpressed recombinantly or chemically synthesized. In some cases, thepeptide is subsequently conjugated to an active agent. The peptide orpeptide-active agent conjugate is administered to a subject in needthereof. The subject is a human or non-human animal. The subject in needthereof is diagnosed with hereditary nephropathy, such as polycystickidney disease or Alport's syndrome. The peptide or peptide-conjugate isdelivered via intravenous administration. Upon administration, thepeptide or peptide conjugate rapidly targets the kidneys, and is used totreat hereditary nephropathy.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 92 Treatment of Interstitial Nephritis

This example describes treatment of interstitial nephritis with thepeptides of the present disclosure. A peptide of this disclosure isexpressed recombinantly or chemically synthesized. In some cases, thepeptide is subsequently conjugated to an active agent. The peptide orpeptide-active agent conjugate is administered to a subject in needthereof. The subject is a human or non-human animal. The subject in needthereof is diagnosed with interstitial nephritis induced by drug use(e.g. Chinese herb induced nephropathy, NSAID induced nephropathy),multiple myeloma, or sarcoid. The peptide or peptide-conjugate isdelivered via intravenous administration. Upon administration, thepeptide or peptide conjugate rapidly targets the kidneys, and is used totreat interstitial nephritis.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 93 Use of Peptides in Renal Transplantation

This example describes the use of peptides of the present disclosure inrenal transplantation. A peptide of this disclosure is expressedrecombinantly or chemically synthesized. In some cases, the peptide issubsequently conjugated to an active agent. The peptide orpeptide-active agent conjugate is administered to a subject in needthereof. The active agent is an anti-rejection drug such as prednisone,azathioprine, mycophenolate mofetil, mycophemolic acid, sirolimius,cyclosporine, or tacrolimus, and the subject is a human or non-humananimal. A donor kidney is needed by the subject, which is treated withthe peptide or peptide conjugate prior to transplantation.Alternatively, the subject is treated post-transplantation for delayedgraft function, acute kidney rejection, or chronic rejection. Forpost-transplantation treatment, the peptide or peptide-conjugate isdelivered via intravenous administration. Upon administration, thepeptide or peptide conjugate rapidly targets the kidneys, and is used totreat post-transplantation kidney conditions.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 94 Use of Peptides to Treat Diabetes or High Blood Pressure

This example describes the use of peptides of the present disclosure totreat diabetes or high blood pressure. A peptide of this disclosure isexpressed recombinantly or chemically synthesized. The peptide isadministered to a subject in need thereof. Ion channels in the kidney(such as sodium channels or potassium channels) are modulated by thepeptide, or the reuptake of glucose is blocked by the peptide. Thesubject is a human or non-human animal. The subject in need thereof isdiagnosed with diabetes or high blood pressure. The peptide is deliveredvia intravenous administration. Upon administration, the peptide rapidlytargets the kidneys and modulates sodium, potassium, or glucosetransport in kidneys and is used to treat diabetes or high bloodpressure.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 95 Use of Peptides to Prevent Renal Fibrosis

This example describes the use of peptides of the present disclosure toprevent renal fibrosis. A peptide of this disclosure is expressedrecombinantly or chemically synthesized. The peptide is conjugated to aplatelet derived growth factor (PDGF) inhibitor. The peptide-drugconjugate is administered to a subject in need thereof. The subject is ahuman or non-human animal. The subject in need thereof is at risk ofrenal fibrosis. The peptide is delivered via intravenous administration.Upon administration, the peptide rapidly targets the kidneys andprevents renal fibrosis.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 96 Oral Delivery to the Kidney

This example describes the oral delivery of peptides of the presentdisclosure. A peptide of this disclosure is expressed recombinantly orchemically synthesized. In some cases, the peptide is subsequentlyconjugated to an active agent. The peptide or peptide-active agentconjugate is administered orally to a subject in need thereof. Thesubject is a human or non-human animal. Upon administration, peptide orpeptide-active agent rapidly targets the kidneys. Optionally, thepeptide is formulated with agents to enhance oral delivery, such aspermeation enhancers such as SNAC, 5-CNAC, sodium caprylate, an aromaticalcohol, EDTA, a sodium alkyl sulfate, or a citrate, or proteaseinhibitors. Some of the peptide is absorbed and traffics to the kidney.The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 97 Peptide Ciprofloxacin Conjugates

This example describes conjugation of peptides of this disclosure tociprofloxacin. A peptide of the disclosure (e.g., any one of SEQ ID NO:1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ IDNO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448,or SEQ ID NO: 451-SEQ ID NO: 529) is expressed recombinantly orchemically synthesized. The peptide is linked to ciprofloxacin via anester link to the carboxylic acid in ciprofloxacin.

The peptide-ciprofloxacin conjugates are administered to a subject. Thesubject can be a human or non-human animal. The subject can have akidney infection (for example, a kidney infection that has spread from aurinary tract infection or a kidney infection that is novel) orpyelonephritis. After administration, the peptide-ciprofloxacinconjugates are homed to the kidneys. Peptide-ciprofloxacin conjugatesare used to prevent loss in kidney function and protect renal functionin subjects with one of the above pre-existing conditions and/or toeliminate or reduce the infection.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 98 Peptide Dapagliflozin Conjugates

This example describes conjugation of peptides of this disclosure todapagliflozin. A peptide of the disclosure (e.g., any one of SEQ ID NO:1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ IDNO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448,or SEQ ID NO: 451-SEQ ID NO: 529) is expressed recombinantly orchemically synthesized. The peptide is linked to dapagliflozin via anester link to the carboxylic acid in dapagliflozin.

The peptide-dapagliflozin conjugates are administered to a subject. Thesubject can be a human or non-human animal. The subject can havediabetes. After administration, the peptide-dapagliflozin conjugates arehomed to the kidneys. Peptide-dapagliflozin conjugates are used toprevent loss in kidney function and protect renal function and/or toimprove glycemic control and reduce damage to other organs due todiabetes by increasing glucose secretion in subjects with one of theabove pre-existing conditions.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 99 Peptide Furosemide Conjugates

This example describes conjugation of peptides of this disclosure tofurosemide. A peptide of the disclosure (e.g., any one of SEQ ID NO:1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ IDNO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448,or SEQ ID NO: 451-SEQ ID NO: 529) is expressed recombinantly orchemically synthesized. The peptide is linked to furosemide via an esterlink to the carboxylic acid in furosemide.

The peptide-furosemide conjugates are administered to a subject. Thesubject can be a human or non-human animal. The subject can havediabetes, hypertension, chronic kidney disease, or nephrotic syndrome.After administration, the peptide-furosemide conjugates are homed to thekidneys. Peptide-furosemide conjugates increase kidney diuresis and areused to prevent loss in kidney function and protect renal function,and/or reduce hypertension and effects thereof in subjects with one ofthe above pre-existing conditions.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 100 Treatment of Renal Insufficiency

This example describes treatment of renal insufficiency with thepeptides of the present disclosure. A peptide of this disclosure isexpressed recombinantly or chemically synthesized. In some cases, thepeptide is subsequently conjugated to an active agent. The peptide orpeptide-active agent conjugate is administered to a subject in needthereof. The subject is a human or non-human animal. The subject in needthereof is diagnosed with renal insufficiency that may be caused byrenal artery disease. The peptide or peptide-conjugate is delivered viaintravenous administration. Upon administration, the peptide or peptideconjugate rapidly targets the kidneys, and is used to treat renalinsufficiency.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570. Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 101 Grafting of Moieties to Enhance Peptide Binding and/orAccumulation in the Kidney

This example describes grafting of other moieties to any peptide of thepresent disclosure (e.g. SEQ ID NO: 33 or SEQ ID NO: 132, SEQ ID NO: 4,SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ ID NO: 1-SEQID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ ID NO:216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448, SEQID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570). Another moiety is graftedto the peptide to enhance binding and/or accumulation in the kidney.Grafting can be performed in a variety of ways. A loop, or fraction of aloop, of a peptide of this disclosure is deleted and replaced with anyof the moieties listed below, or a fraction thereof. Such a peptide ofthis disclosure is expressed recombinantly or chemically synthesized.Or, grafting is done by inserting the nucleic acid sequence of the othermoiety into the vector comprising a nucleic acid sequence of a peptideof the present, which is then expressed as a fusion of the other moietyand the peptide of the present disclosure. Alternatively, the othermoiety is conjugated to a recombinantly expressed or chemicallysynthesized peptide of this disclosure. Other moieties that are graftedor conjugated to any peptide of this disclosure include Y(KKEEE)₃K (SEQID NO: 624), Y(KKEE)₅K (SEQ ID NO: 625), Y(KKQQQ)₃K (SEQ ID NO: 626),Y(MARIA)₃ (SEQ ID NO: 627), (KKEEE)₃K (SEQ ID NO: 628), (KKEE)₅K (SEQ IDNO: 629), (KKQQQ)₃K (SEQ ID NO: 630), (MARIA)₃ (SEQ ID NO: 631),(APASLYN)₂ (SEQ ID NO: 632), and ANTPCGPYTHDCPCKR (SEQ ID NO: 633).(Janzer et al. Bioconjug Chem. Oct. 4, 2016, Geng et al. Bioconjug Chem.Jun. 20, 2012; 23(6):1200-10, Wischnjow et al. Bioconjug Chem. Apr. 20,2016; 27(4):1050-7). Any L-Tyr residue in any of the foregoing can bemodified to D-Tyr, for example, for the purposes of radiolabeling.

Grafting of these other moieties to a peptide of this disclosure canconfer additional targeting properties by enhancing, changing, ormodifying the properties of the peptides of the present disclosure.Other moieties contain positively charged residues, which increasingbinding of peptides to proximal tubule cells, to megalin (which isnegatively charged), or otherwise increase retention in the kidney.Other moieties also modify the properties of the peptides of thisdisclosure by changing charge, changing absorption properties into theproximal tubules, or changing targeting of specific structures withinthe kidney.

Example 102 Peptide Torsemide Conjugates

This example describes conjugation of peptides of this disclosure totorsemide. A peptide of the disclosure (e.g., any one of SEQ ID NO:1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQ IDNO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO: 448,or SEQ ID NO: 451-SEQ ID NO: 529) is expressed recombinantly orchemically synthesized. The peptide is linked to torsemide via acleavable or stable linker.

The peptide-torsemide conjugates are administered to a subject. Thesubject can be a human or non-human animal. The subject can have edemaas a result of kidney disease or high blood pressure. Afteradministration, the peptide-torsemide conjugates are homed to thekidneys. Peptide-torsemide conjugates increase kidney diuresis and areused to prevent loss in kidney function and protect renal function,and/or reduce hypertension and effects thereof in subjects with one ofthe above pre-existing conditions.

The peptide can be a peptide of SEQ ID NO: 33, SEQ ID NO: 132, SEQ IDNO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ ID NO: 6, or any of SEQ IDNO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO: 206, SEQ ID NO: 213, SEQID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQ ID NO: 441, SEQ ID NO:448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO: 570 Such peptide-drugconjugates can be made using either a cleavable or stable linker asdescribed herein (e.g., EXAMPLES 14 and 15).

Example 103 Fluorescence of Kidney Homing Peptides

This example illustrates peptide homing to kidney after administrationof a peptide fluorophore conjugate. A peptide of any one of SEQ ID NO:33, SEQ ID NO: 132, SEQ ID NO: 4, SEQ ID NO: 41, SEQ ID NO: 5, or SEQ IDNO: 6, or any of SEQ ID NO: 1-SEQ ID NO: 120, SEQ ID NO: 127-SEQ ID NO:206, SEQ ID NO: 213, SEQ ID NO: 216-SEQ ID NO: 355, SEQ ID NO: 362-SEQID NO: 441, SEQ ID NO: 448, SEQ ID NO: 451-SEQ ID NO: 529, or SEQ ID NO:570 is chemically conjugated to a cyanine, tricarboxocyanine or otherfluorescent dye and then imaged using, for example, the methods ofEXAMPLE 59.

Example 104 Peptide Resistance Under Various Conditions

This example illustrates peptide stability under various stressconditions such as high temperature, low pH, reducing agents, andproteases. To determine resistance to high temperatures, cystine-densepeptides (CDPs) are incubated at 0.5 mM in PBS at 75° C. or 100° C. for1 h and pelleted, and the supernatant is analyzed with reversed-phasechromatography (RPC). To determine resistance to proteolytic digestion,CDPs are mixed with 50 U of porcine pepsin, in simulated gastric fluidat pH 1.0, or 50 U of porcine trypsin in PBS, incubated for 30 minutesat 37° C. and analyzed with RPC. Oxidized and reduced forms (preparedthrough addition 10 mM DTT) are compared. Circular Dichroismspectroscopy is used in order to measure the secondary structure ofpeptides with a Jasco J-720W spectropolarimeter in a cell with a 1.0-mmpath length, and CDPs are diluted into 20 mM phosphate buffer, pH 7.4,at a concentration of 15-25 μM. These conditions are expected todenature or degrade conventional globular proteins and many peptides. Inscoring the results, “high” resistance indicates a high amount of thepeptide remains or is retained as unmodified under the givenexperimental conditions and “low” resistance indicates a low amount ofthe peptide remains or is retained unmodified under the givenexperimental conditions. Notably, the experimental conditions describedin this example are more extreme stress conditions than many standard invivo or physiologic conditions, in vitro conditions, conditions duringmanufacturing, and handling conditions. As such, even “low” resistancein this assay can indicate meaningful resistance to these stressconditions that may have applicability for a number of uses describedherein.

While certain embodiments of the present disclosure have beenexemplified or shown and described herein, it will be apparent to thoseskilled in the art that such embodiments are provided by way of exampleonly. It is not intended that the disclosure be limited by the specificexamples provided within the specification. While the disclosure hasbeen described with reference to the aforementioned specification, thedescriptions and illustrations of the embodiments herein are not meantto be construed in a limiting sense. Numerous variations, changes, andsubstitutions will now occur to those skilled in the art withoutdeparting from the disclosure. Furthermore, it shall be understood thatall embodiments of the disclosure are not limited to the specificdepictions, configurations or relative proportions set forth hereinwhich depend upon a variety of conditions and variables. It should beunderstood that various alternatives to the embodiments of thedisclosure described herein may be employed in practicing thedisclosure. It is therefore contemplated that the disclosure shall alsocover any such alternatives, modifications, variations or equivalents.It is intended that the following claims define the scope of thedisclosure and that methods and structures within the scope of theseclaims and their equivalents be covered thereby.

What is claimed:
 1. A peptide active agent conjugate, the peptide activeagent conjugate comprising: a) a peptide comprising a sequence that has:(i) at least 80% sequence identity to SEQ ID NO: 240 or a fragmentthereof that has at least 90% sequence identity to at least 19 aminoacid residues of SEQ ID NO: 240, or (ii) at least 80% sequence identityto SEQ ID NO: 239 or a fragment thereof that has at least 90% sequenceidentity to at least 19 amino acid residues of SEQ ID NO: 239, at least80% sequence identity to SEQ ID NO: 268 or a fragment thereof that hasat least 90% sequence identity to at least 15 amino acid residues of SEQID NO: 268, at least 80% sequence identity to SEQ ID NO: 276 or afragment thereof that has at least 90% sequence identity to at least 8amino acid residues of SEQ ID NO: 276, or at least 95% sequence identityto SEQ ID NO: 241 or a fragment thereof that has at least 95% sequenceidentity to at least 15 amino acid residues of SEQ ID NO: 241; and b) anactive agent, wherein the active agent is secukinumab, brodalumab,ixekizumab, tocilizumab, sarilumab, ALX-0061, sirukumab, clazakizumab,olokizumab, MEDI5117, ustekinumab, briakinumab, tildrakizumab,guselkumab, soluble IL-23, cytokine-binding homology region of solubleIL-23, canakinumab, rilonacept, gevokizumab, LY2189102, anakinra,MEDI-8968, AMG-108, kineret, type I interferon, or IFN-beta.
 2. Thepeptide active agent conjugate of claim 1, wherein the active agent issecukinumab, ustekinumab, tocilizumab, anakinra, or kineret.
 3. Thepeptide active agent conjugate of claim 1, wherein the peptide comprises4 or more cysteine residues.
 4. The peptide active agent conjugate ofclaim 1, wherein the peptide or the fragment thereof comprises three ormore disulfide bridges formed between cysteine residues, wherein one ofthe disulfide bridges passes through a loop formed by two otherdisulfide bridges.
 5. The peptide active agent conjugate of claim 1,wherein the peptide or the fragment thereof comprises 5 to 12 basicresidues.
 6. The peptide active agent conjugate of claim 1, wherein thepeptide or the fragment thereof comprises from 0 to 5 acidic residues.7. The peptide active agent conjugate of claim 1, wherein the peptidehas a charge greater than 2 at physiological pH.
 8. The peptide activeagent conjugate of claim 1, wherein 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10active agents are linked to the peptide.
 9. The peptide active agentconjugate of claim 1, wherein the peptide is SEQ ID NO: 240, SEQ ID NO:239, SEQ ID NO: 268, SEQ ID NO: 276, or SEQ ID NO:
 241. 10. The peptideactive agent conjugate of claim 1, further comprising a linkage betweenan N-terminus or a C-terminus of the peptide and the active agent. 11.The peptide active agent conjugate of claim 1, further comprising alinker linking the peptide to the active agent at an N-terminus, at theepsilon amine of an internal lysine residue, at the carboxylic acid ofan aspartic acid or glutamic acid residue, or a C-terminus of thepeptide.
 12. The peptide active agent conjugate of claim 1, wherein thepeptide comprises a non-natural amino acid, wherein the non-naturalamino acid is an insertion, appendage, or substitution for another aminoacid, and wherein the peptide conjugate comprises a linkage between thenon-natural amino acid and the active agent.
 13. The peptide activeagent conjugate of claim 1, further comprising a cleavable linkerlinking the peptide to the active agent, wherein the cleavable linkercomprises a cleavage site for matrix metalloproteinases, thrombin,cathepsins, or beta-glucuronidase.
 14. The peptide active agentconjugate of claim 1, further comprising a linker linking the peptide tothe active agent, wherein the linker comprises an amide bond, an esterbond, a carbamate bond, a carbonate bond, a hydrazone bond, an oximebond, a disulfide bond, a thioester bond, a thioether bond, a triazole,a carbon-carbon bond, or a carbon-nitrogen bond.
 15. The peptide activeagent conjugate of claim 1, further comprising a hydrolytically labilelinker linking the peptide to the active agent.
 16. The peptide activeagent conjugate of claim 1, further comprising a linker linking thepeptide to the active agent, wherein the linker is pH sensitive,reducible, glutathione-sensitive, or protease cleavable.
 17. The peptideactive agent conjugate of claim 1, further comprising a stable linkerlinking the peptide to the active agent.
 18. The peptide active agentconjugate of claim 1, further comprising an Fc domain of an antibodylinking the peptide to the active agent.
 19. A pharmaceuticalcomposition comprising the peptide active agent conjugate of claim 1, ora salt thereof, and a pharmaceutically acceptable carrier.
 20. A methodof treating a kidney disorder in a subject comprising administering tothe subject a peptide active agent conjugate comprising: a. a peptidecomprising a sequence that has: (i) at least 80% sequence identity toSEQ ID NO: 240 or a fragment thereof that has at least 90% sequenceidentity to at least 19 amino acid residues of SEQ ID NO: 240, or (ii)at least 80% sequence identity to SEQ ID NO: 239 or a fragment thereofthat has at least 90% sequence identity to at least 19 amino acidresidues of SEQ ID NO: 239, at least 80% sequence identity to SEQ ID NO:268 or a fragment thereof that has at least 90% sequence identity to atleast 15 amino acid residues of SEQ ID NO: 268, at least 80% sequenceidentity to SEQ ID NO: 276 or a fragment thereof that has at least 90%sequence identity to at least 8 amino acid residues of SEQ ID NO: 276,or at least 95% sequence identity to SEQ ID NO: 241 or a fragmentthereof that has at least 90% sequence identity to at least 15 aminoacid residues of SEQ ID NO: 241; and b. an active agent, wherein theactive agent is secukinumab, brodalumab, ixekizumab, tocilizumab,sarilumab, ALX-0061, sirukumab, clazakizumab, olokizumab, MEDI5117,ustekinumab, briakinumab, tildrakizumab, guselkumab, soluble IL-23,cytokine-binding homology region of soluble IL-23, canakinumab,rilonacept, gevokizumab, LY2189102, anakinra, MEDI-8968, AMG-108,kineret, type I interferon, or IFN-beta; thereby treating the kidneydisorder.
 21. The method of claim 20, wherein the disorder is chronickidney disease, lupus nephritis, diabetic nephropathy, renal fibrosis,acute kidney injury, chronic glomerulonephritis, interstitial nephritis,kidney transplantation, high blood pressure, or hypertensivenephrosclerosis.
 22. The method of claim 20, wherein upon administrationto the subject the peptide active agent conjugate homes, targets,migrates to, accumulates in, binds to, is retained by, or is directed toa kidney of the subject.
 23. The method of claim 20, wherein uponadministration to the subject, the peptide active agent conjugate homesto proximal tubules of a kidney.
 24. The method of claim 20, wherein theadministering comprises administering by inhalation, intranasally,orally, topically, parenterally, intravenously, subcutaneously,intra-articularly, intramuscularly administration, intraperitoneally,dermally, transdermally, or a combination thereof.
 25. A method oflabeling a renal tissue of a subject comprising administering to thesubject a peptide active agent conjugate comprising: a. a peptidecomprising a sequence that has: (i) at least 80% sequence identity toSEQ ID NO: 240 or a fragment thereof that has at least 90% sequenceidentity to at least 19 amino acid residues of SEQ ID NO: 240, or (ii)at least 80% sequence identity to SEQ ID NO: 239 or a fragment thereofthat has at least 90% sequence identity to at least 19 amino acidresidues of SEQ ID NO: 239, at least 80% sequence identity to SEQ ID NO:268 or a fragment thereof that has at least 90% sequence identity to atleast 15 amino acid residues of SEQ ID NO: 268, at least 80% sequenceidentity to SEQ ID NO: 276 or a fragment thereof that has at least 90%sequence identity to at least 8 amino acid residues of SEQ ID NO: 276,or at least 95% sequence identity to SEQ ID NO: 241 or a fragmentthereof that has at least 95% sequence identity to at least 15 aminoacid residues of SEQ ID NO: 241; b. an active agent, wherein the activeagent is secukinumab, brodalumab, ixekizumab, tocilizumab, sarilumab,ALX-0061, sirukumab, clazakizumab, olokizumab, MEDI5117, ustekinumab,briakinumab, tildrakizumab, guselkumab, soluble IL-23, cytokine-bindinghomology region of soluble IL-23, canakinumab, rilonacept, gevokizumab,LY2189102, anakinra, MEDI-8968, AMG-108, kineret, type I interferon, orIFN-beta; and c. a detectable agent; and homing the peptide active agentconjugate to the renal tissue, thereby labeling the renal tissue. 26.The peptide active agent conjugate of claim 25, wherein the detectableagent is a fluorophore, a near-infrared dye, a contrast agent, ananoparticle, a metal-containing nanoparticle, a metal chelate, an X-raycontrast agent, a PET agent, a radioisotope, or a radionuclide chelator.27. The method of claim 25, further comprising imaging the renal tissueand detecting a cancer or diseased region, tissue, or cell based on thelabeling.
 28. The method of claim 27, further comprising performingsurgery on the subject and removing the cancer or diseased region,tissue, or cell.
 29. A peptide active agent conjugate, the peptideactive agent conjugate comprising: a. a peptide comprising a sequencethat has: (i) at least 80% sequence identity to SEQ ID NO: 240 or afragment thereof that has at least 90% sequence identity to at least 19amino acid residues of SEQ ID NO: 240, or (ii) at least 80% sequenceidentity to SEQ ID NO: 239 or a fragment thereof that has at least 90%sequence identity to at least 19 amino acid residues of SEQ ID NO: 239,at least 80% sequence identity to SEQ ID NO: 268 or a fragment thereofthat has at least 90% sequence identity to at least 15 amino acidresidues of SEQ ID NO: 268, at least 80% sequence identity to SEQ ID NO:276 or a fragment thereof that has at least 90% sequence identity to atleast 8 amino acid residues of SEQ ID NO: 276, or at least 95% sequenceidentity to SEQ ID NO: 241 or a fragment thereof that has at least 95%sequence identity to at least 15 amino acid residues of SEQ ID NO: 241;and b. an active agent, wherein the active agent is resveratrol,arglabin, desciclesonide, iguratimod, montelukast, rofecoxib,valdecoxib, MCC950, or β-hydroxybutyrate (BHB).
 30. The peptide activeagent conjugate of claim 29, wherein the peptide is SEQ ID NO: 240, SEQID NO: 239, SEQ ID NO: 268, SEQ ID NO: 276, or SEQ ID NO: 241.